Recent advances in S-palmitoylation and its emerging roles in human diseases

Abstract

S-palmitoylation, one reversible post-translational modification (PTM) involving the binding of palmitate to proteins, influences protein stability, localization and interactions. Through S-palmitoylation, proteins can be targeted to specific cellular compartments, form functional complexes, and participate in intricate signaling cascades. Organized and reversible S-palmitoylation process achieves remarkable roles in the precise orchestration of biological activities, including cell signaling, membrane trafficking, synaptic transmission and cellular immunity. S-palmitoylation has been implicated in the pathogenesis of diverse disorders including cancers, cardiovascular diseases, metabolic diseases, immunological diseases, infection diseases, nervous system and mental diseases. Altered S-palmitoylation of proteins changes the oncogenic function, synaptic localization, enzymatic activity and signaling transduction, potentially contributing to disease progression. Understanding and targeting S-palmitoylation pathways hold promise for therapeutic interventions in associated diseases. Collectively, S-palmitoylation is a key regulatory mechanism with significant implications for disease pathogenesis. Investigating the role of S-palmitoylation provides insights into diagnostic markers and potential therapeutic targets, highlighting the importance of ongoing research in understanding the broader implications of S-palmitoylation in health and disease.

Introduction

There are over 1 million proteins in human proteome encoded by only 20,000 to 25,000 genes [1], which rely on complex mechanisms including alternative splicing, co-translational and post-translational modifications (PTMs). PTMs occur at amino acids by modifying with multiple chemical additions during or after protein biosynthesis, such as methyl, phosphoryl, glycosyl and acetyl groups, conferring specific functions to proteins [2–4]. To date, the UniProt database has listed more than 600 types of PTMs, which play a vital role in protein localization, secretion, and function [5, 6].

Palmitoylation is the most common lipid acylation modification and affects about 20% of the proteome in mammals [7]. This lipid modification involves the addition of palmitate, a 16-carbon saturated fatty acid (FA), to specific cysteine residues of proteins, thereby modulating their properties and activities. Depending on different binding sites, palmitoylation can be classified into N-, O-, and S-palmitoylation [8, 9]. S-palmitoylation, also referred to S-acylation, is the predominant and only reversible form [10, 11]. This dynamic nature grants S-palmitoylation the versatility to fine-tune protein functions in response to diverse cellular signaling and environmental stimuli. The balance between S-palmitoylation and depalmitoylation is essential in many biological processes, including cell signaling, membrane trafficking, synaptic transmission, as well as immune responses. Increasing evidence has revealed that abnormalities in S-palmitoylation and depalmitoylation is associated with a range of human disorders, such as cancers, cardiovascular diseases, metabolic diseases, immunological diseases, nervous system and mental diseases [12–14].

In the past decades, Maurine E Linder had reviewed the addition of palmitate to membrane proteins [11]. There are also reviews about S-palmitoylation in specific pathology, including inflammation [15], cancers [16, 17]. In this context, researchers need to elucidate the mechanism governing S-palmitoylation, identify S-palmitoylated proteins, and decipher the functional consequence of this modification to further understand cellular biology and disease processes. The exploration of S-palmitoylation represents a frontier in molecular research with far-reaching implications for both basic science and clinical applications. In this review, we summarize the current understanding of the S-palmitoylation and depalmitoylation machinery, highlight its roles in various human diseases, and discuss emerging therapeutic strategies targeting this versatile PTM.

The process and regulation of S-palmitoylation

S-palmitoylation refers to the covalent binding of a palmitoyl group to cysteine residues of proteins. It is the only reversible form of palmitoylation because thioester bonds can be hydrolyzed under certain conditions [11]. The source of palmitate is closely tied to cellular metabolism. (Fig. 1). Glucose is transported by glucose transporter (GLUT) and converted to generate pyruvate via glycolytic pathway [18]. Within mitochondria, pyruvate is catalyzed by pyruvate dehydrogenase to produce acetyl-coenzyme A (acetyl-CoA). Acetyl-CoA is transformed into malonyl-CoA through the action of acetyl-CoA carboxylase (ACC), and then FA synthase (FASN) catalyzes them to form palmitic acid (PA). Palmitate can be also imported into cell across the plasma membrane. Palmitate in the cytoplasm is then catalyzed by Acyl-CoA synthetase (ACS) to form palmitoyl-CoA [19]. S-palmitoylation is primarily catalyzed by a family of enzymes known as palmitoyl acyltransferases (PATs), also referred to as ZDHHC enzymes, named for their conserved Asp-His-His-Cys (DHHC) zinc-finger motif. The process of S-palmitoylation occurs in two main steps: (1) ZDHHC auto-palmitoylation. The thiolate nucleophile of Cys156 attacks carbonyl carbon of palmitoyl-CoA thioester to form a transfer intermediate, which could link palmitoyl-CoA to Cys156 in ZDHHC domain [20, 21]. Palmitoyl-CoA partitions into the lipid bilayer and likely distorts the local structure of the membrane, where interfaces of membrane proteins with surrounding lipids are accumulated. Palmitoyl-CoA inserts into the cavity formed by His154 and aspartic acid in ZDHHC motif from within the membrane, which can accept a proton from ZDHHC cysteine, enabling nucleophilic attack on the carbonyl thioester of palmitoyl-CoA [20]. (2) Substrate palmitoylation. The activated palmitoyl group is then transferred from the ZDHHC enzyme to a cysteine residue on the target protein via a thioester bond [22]. Protonated His154 activates the acyl-enzyme thioester for the subsequent palmitoyl transfer to the substrate by providing a proton to the carbonyl oxygen [21]. S-palmitoylation can be reversed by serine hydrolase superfamily, such as acyl-protein thioesterases (APTs) and palmitoyl-protein thioesterases (PPTs) [23, 24]. This dynamic cycle of S-palmitoylation and depalmitoylation occurs from seconds to hours, enabling proteins to rapidly traffic among specific cellular areas and efficiently catalyzed biological reactions [25, 26].

Fig. 1.

The process and regulation of S-palmitoylation cycle. S-palmitoylation refers to palmitoyl group covalently binds to cysteine residues of protein, mainly catalyzed by PATs. S-palmitoylation can be reversed by serine hydrolase superfamily, such as APTs. Abbreviations: ACC, acetyl-CoA carboxylase; ACS, acyl-CoA synthetase; APTs, acyl-protein thioesterases; CMA, cyanomethyl-myracrylamide; FASN, fatty acid synthase; Glu, glucose; GLUT, glucose transporter; PA, palmitic acid; PATs, protein acyltransferases; 2-BP, 2-bromopalmitate

ZDHHC enzymes

In recent years, 23 kinds of ZDHHC enzymes have been reported in the human proteome (ZDHHC1-24, excluding ZDHHC10) (Table 1), which are widely localized in membrane-bound organelles, including the endoplasmic reticulum (ER), Golgi apparatus (GA), and mitochondria [27, 28]. ZDHHC enzymes share a common membrane topology characterized by at least 4 transmembrane domains (TMD). DHHC cysteine-rich domain is localized within an intracellular loop between TMD2 and TMD3, which confines S-palmitoylation to occur at the membrane-cytosol interface (Fig. 2a) [20, 29]. ZDHHCs also possess other conserved motifs including palmitoyl transferase conserved carboxy-terminal (PaCCT), a threonine-threonine-x-glutamic acid (TTxE) and aspartic acid-proline-glycine (DPG) (Fig. 2a) [29]. The DHHC domain is rich in cysteine, which endows ZDHHC enzyme with catalytic activity. Some enzymes have high selectivity, while others are relatively promiscuous (Table 1) [28, 30, 31]. For example, gasdermin family proteins GSDME, which widely participate in cytokine secretion and pyroptosis, can be S-palmitoylated by ZDHHC2, 7, 11, 15 [32]. More than half of ZDHHCs is observed to be capable of increasing interferon-induced transmembrane protein-3 (IFITM3) S-palmitoylation, but only ZDHHC20 shows extensive colocalization with IFITM3 at lysosomes and uniquely enhances its antiviral activity [33].

Table 1.

The biological features and inhibitors of S-palmitoylation, depalmitoylation enzymes

Enzyme	Genomic location [1]	Localization	Interaction domains	Substrates	Inhibitors [2]
S-palmitoylation enzymes
ZDHHC1	Chr16: 67,394,152 − 67,416,833	ER		NLRP3 [197], IFITM3 [198], IGF2BP1 [199], p53 [200], Gpm6a [201]
ZDHHC2	Chr8: 17,156,482 − 17,224,799	plasm membrane, endosome		AGK [202], GSDME [32], Gpm6a [201], HA [203], nsP1 [204], AKAP150 [205]	TTZ-1, TTZ-2 [301]
ZDHHC3	Chr3: 44,915,257 − 44,976,185	Golgi	C157	Cadm4 [206], PD-L1 [95], GSDME [207], IRHOM2 [141], Rac1 [208], SLC9A2 [209], GluA1 [210], VMP1 [211], PML/RARα [129], ACE2 [212], SCAP [213], JAK [214], PRCD [215], SARS-CoV-2 E [216], NCAM [40], D2R [217]	circPDIA3 [207]
ZDHHC4	Chr7: 6,577,434-6,589,374	ER		CD82 [218], MAVS [219], CD36 [142], TRPV1 [220], GSK3β [221], NFATC4 [68], D2R [217]
ZDHHC5	Chr11: 57,667,747 − 57,701,190	plasm membrane	PDZ-binding motif	GRIP1b [34], PSD95 [222], CD36 [142], GSDMD [223], NOD1/2 [151], CLOCK [224], NLRP3 [225], PKCδ [226], lamin A [227], Furin/PC7 [228], PCDH7 [229], Fz5 [230], PLM [231], NCX1 [45], EZH2 [39], SSTR5 [232], SMPDL3B [233], STAT3 [234], TrpM7 [235], Gp130 [236]	Artemether [226], Lomitapide [232]
ZDHHC6	Chr10: 112,424,412 − 112,447,572	ER	SH3-2 domain	multiple type I and II membrane proteins [237], SARS-CoV-2 E [216], FLT3-ITD [125], CLIMP-63 [238], MYD88 [239], AEG-1 [240], PPARγ [241], Nras [242], DGKε [243]	Artemisinin [242]
ZDHHC7	Chr16: 85,007,781 − 85,045,141	Golgi		GSDME [32], HA [203], Rac1 [208], DGKε [243], STAT3 [58], ATG16L1 [185], NLRP3 [244], Septin 8 [245], KLF10 [246], SCRIB [247], MAVS [248], JAK [214], NCAM [40], APP [249], APT2 [59]	MY-D-2-4-5-6, Cadmium chloride, NCI 68,093, Zinc pyrithione
ZDHHC8	Chr22: 20,129,456 − 20,148,007	Golgi, plasm membrane	PDZ-binding motif	GRIP1b [34], NFATC4 [68], SLC7A11 [41], Gp130 [236], PSD95 [250], D2R [217]	Lithium
ZDHHC9	ChrX: 129,803,288 − 129,843,909	Golgi, ER		GSDMD [223], cGAS [251], LDHA [252], Bip protein [118], Rab3gap1 [253], APLNR [254], β2AR [255]	Artemisinin, Bis-piperazine backbone compounds 13 and 25
ZDHHC11	Chr5: 795,605–860,563	plasm membrane		GSDME [32], ATGL [256], the envelope protein of Zika virus [257]
ZDHHC12	Chr9: 128,720,870 − 128,724,127	Golgi, ER		SARS-CoV-2 E [216], CLDN3 [258], NLRP3 [119]
ZDHHC13	Chr11: 19,117,099 − 19,176,422	Golgi, ER	ankyrin repeat domains	SNAP25b [259], HTT [30], JNK [260], CSP [51], MC1R [261], ULK1 [262], EGFR [263], Drp1 [264], loricrin, peptidyl arginine deiminase type III, keratin fiber crosslinker transglutaminase 1 [265], cornifelin [266]
ZDHHC14	Chr6: 157,381,133–157,678,157	Golgi, plasm membrane		PSD93 [36], GSDMD [137]
ZDHHC15	ChrX: 75,368,427 − 75,523,502	Golgi		GSDME [32], HA [203], SARS-CoV-2 E [216], GP130 [236], Nrp2 [267]	TTZ1, TTZ2
ZDHHC16	Chr10: 97,446,170 − 97,457,370	ER		DGKε [243], PCSK9 [111], SETD2 [268]
ZDHHC17	Chr12: 76,764,103 − 76,853,701	Golgi	ankyrin repeat domains	SNAP25b [259], HTT [30], JNK [260], CSP [51], GSDME [207], TrpM7 [235], DGKε [243], AKT [269], Oct4A [270], DLK, NMNAT2 [271], SPRED3 [272], NLRP3 [273], Sept8-204 [245], Smad7 [274], Sprouty-2 [275], ankyrin-B [276], NMNAT2 [277]	Metoprolol, Minocycline, Fluorodeoxyglucose
ZDHHC18	Chr1: 26,826,688 − 26,857,604	Golgi		MDH2 [278], cGAS [279]	MY-D-4
ZDHHC19	Chr3: 196,197,452 − 196,211,437	ER		nsP1 [204], STAT3 [280], SQSTM1 [281], p62 [282], Flot-1 [283], Smad3 [99]
ZDHHC20	Chr13: 21,348,784 − 21,459,370	plasm membrane		HA [203], SARS-CoV-2 E [216], YTHDF3 [114], CD80 [284], MBLAC2 [285], ORAI1 [286], IFITM3 [33]
ZDHHC21	Chr9: 14,586,766 − 14,693,432	plasm membrane, Golgi		AK2 [128], 5-HT1AR [175], FASN [133], TRPV2 [287], Cav-2 [288], α1-adrenergic receptor [289]
ZDHHC22	Chr14: 77,131,270 − 77,142,734	ER, Golgi		mTOR [290], BK [291], CCN3 [292]
ZDHHC23	Chr3:113,947,901 − 113,979,508	nucleus, vesicle		PHF2 [293], GFAP [294], BK [291]
ZDHHC24	Chr11: 66,520,625 − 66,546,235	plasm membrane, ER, Golgi		AKT [269], MAVS [295]
Depalmitoylation enzymes
APT1	Chr8: 54,042,987 − 54,102,017	mitochondrion, plasm membrane		G protein α subunit [293], the proto-oncogene H-RAS product, SNAP-23, β2-adrenergic receptor [9]	ML348, ML221, palmostatin B and M
APT2	Chr1: 23,791,110 − 23,795,539	Golgi		DHHC6, Growth-associated protein-43 [294], TNF receptor, Melanocortin receptor [9]	ML349, ML221, palmostatin B and M
ABHD8	Chr19: 17,292,131 − 17,310,236			NLRP3 [62]
ABHD10	Chr3: 111,979,010–111,993,368	mitochondrion		PRDX5 [63]
ABHD16A	Chr6: 31,686,955 − 31,703,356	ER		IFITM [295]	C7600, 12-thiazole abietanes [302]
ABHD17A	Chr19: 1,876,809-1,885,495			PSD95 [65], MAP6 [66]
ABHD17B	Chr9: 71,862,452 − 71,911,193	vesicle		HK1 [61], PSD95 [65], N-Ras [110], MAP6 [66]	Palmostatin B, ABD957
ABHD17C	Chr15: 80,679,684 − 80,755,621			PSD95 [65], MAP6 [66]
PPT1	Chr1: 40,071,461 − 40,097,260	lysosome		TLR9 [151], AEG-1 [240], NLRP3 [296], Gpx1 [297], Septin 8 [245], GFAP [298], CSPα [299], AMPAR [300]	Palmostatin B, GNS561 [303], DC661 [304], HDSF, Lucanthone [305], Ezurpimtrostat [306]

^[1] Genomic location refers to GeneCards (https://www.genecards.org/), till July 2025

^[2] 2-bromopalmitate (2-BP) and cyanomethyl-myracrylamide (CMA) are omitted in this table, because they are non-selective

Fig. 2.

ZDHHCs and depalmitoylation enzymes. (a) Canonical topology of ZDHHC enzyme. ZDHHC enzymes share a common membrane topology characterized by at least 4 TMDs, and DHHC cysteine-rich domain is localized within an intracellular loop between TMD2 and TMD3. ZDHHCs also possess other conserved motifs including PaCCT, TTxE and DPG. (b) Phylogenetic tree, amino acid counts, and conserved binding domains of ZDHHCs. (c) Amino acid counts of depalmitoylation enzymes. (d) Topology of canonical ABHD and APT1. ABHD has eight β strands (β1–8) and five α helices (αA-F). Compared to ABHD, APT1 has seven β strands (β2–8, the canonical strand 1 is missing), five α helices (αA-F), four short β irregular strands (S1-4), and four helical segments (G1-4). Abbreviations: ABDHs, α/β hydrolase domain-containing proteins; APT, acyl-protein thioesterase; DHHC, aspartate-histidine-histidine-cysteine; DPG, aspartic acid-proline-glycine; PaCCT, palmitoyl transferase conserved carboxy-terminal; PPT, palmitoyl-protein thioesterase; TMD, transmembrane domain; TTxE, threonine-threonine-x-glutamic acid; ZDHHC, zinc-finger and conserved DHHC domain

Due to the functional redundancy of ZDHHC and substrate redundancy complicates, the recognition mechanism of ZDHHC still remains unclear. Subcellular distribution provides a preliminary level of specificity, because it dictates where ZDHHCs and substrates can interact [7]. Some unique protein-protein interaction motifs, such as ankyrin repeats, SH3 domain, and PDZ binding domain, may provide docking capabilities for substrate selectivity (Fig. 2b) [34, 35]. For example, glutamate receptor interacting protein-1 (GRIP1b), one multi-PDZ domain-containing protein, is S-palmitoylation by ZDHHC5/8, relying on a PDZ ligand-dependent recognition mechanism [35, 36]. The fusion of ankyrin repeats to the N-terminus of ZDHHC3 endows its ability to regulate S-palmitoylation of huntingtin proteins, which would otherwise not occur [37]. Besides, the conserved motifs including PaCCT, TTxE and DPG, have been reported to be potentially involved in substrate protein recognition through certain contacts with the ZDHHC domain. The second threonine of TTxE motif could form a hydrogen bond via directly interacting with aspartic acid in ZDHHC, and Asn266 in PaCCT motif assists side chain in forming an amide via exploiting its H-bond [20].

The stability, localization, and catalytic activity of ZDHHCs rely on transcriptional regulation, PTMs, accessory factors, and substrate concentrations. A recent study revealed that virus infection could change the transcriptional initiation site of ZDHHC20 gene, presenting the first evidence for ZDHHC regulation in transcriptional level [38]. In glioma, frameshift and missense mutations of p53 enhance the interaction of it with NF-γ, both of them are then recruited onto ZDHHC5 promoter (1 kb upstream of it contains four potential NF-γ binding sites). This alteration upregulates enhancer of zeste homolog (EZH2) palmitoylation at Cys571 and Cys576 sites, contributing to the maintenance of glioma cell stemness [39]. PTMs influence ZDHHCs functional activities and properties. For example, fibroblast growth factor receptor (FGFR) and Src proteins phosphorylate ZDHHC3 to modulate its enzymatic activity. Site-directed mutagenesis found that Tyr18 of ZDHHC3 is a FGFR-specific phosphorylation site, while Src kinase catalyzes the phosphorylation of Tyr295 and Tyr297 sites. Abrogation of tyrosine phosphorylation of ZDHHC3 could increase its auto-palmitoylation and enhance the enzymatic activity [40]. Likewise, adenosine activated protein kinase alpha-1 (AMPKα1)-mediated phosphorylation of ZDHHC8 is required for its interaction with SLC7A11 [41]. ZDHHC enzymes themselves undergo S-palmitoylation at their catalytic cysteine site, which is auto-palmitoylated prior to substrate modification. For instance, C-terminal tail of ZDHHC5 is found to be S-palmitoylated, and then mediates response to subsequent molecules [42–44]. Jun Noritake et al. proposed that extracellular signals translocate specific ZDHHC and create route for substrate shuttling. Dendritically localized ZDHHC2 could sense synaptic changes and rapidly translocate near postsynaptic membranes, contributing to rapid palmitoylation and synaptic accumulation of postsynaptic density protein-95 (PSD95) [45].

Accessory factors enable ZDHHCs to participate in a variety of biological processes. For example, ZDHHC9 requires a necessary co-factor GCP16 to stabilize the structure of heterodimers, enabling ZDHHC9 to adopt a conformation that is capable of efficient catalysis. GCP16 also assists ZDHHC14 and 18 [46, 47]. Golgin A7 family member B (Golga7b) is a related isoform of GCP16, which was found to influence the trafficking and localization of ZDHHC5 in non-neuronal cells. Published studies had found that Golga7b imparted a reciprocal regulation on ZDHHC5 via stabilizing the enzyme at plasm membrane. An assessment of endocytosis showed that Golga7b expression reduced the internalization of ZDHHC5, whereas S-palmitoylation-deficient mutant Golga7b upregulated endocytosis of ZDHHC5 [48, 49]. Similarly, ZDHHC6 requires Selenoprotein K (SELENOK) to help facilitate addition of palmitate to cysteine [50]. Kun Huang et al. found that S-palmitoylation of synaptosome associated protein-25 (SNAP25) by ZDHHC17 was potentiated in vitro with the presence of wild-type huntingtin (HTT) protein, which acted as a positive modulator of the S-palmitoylation activity of ZDHHC17 [51]. Interestingly, ZDHHCs have been reported to undergo S-palmitoylation cascades. ZDHHC6 is S-palmitoylated based on upstream ZDHHC16, which acts as PAT, and ZDHHC16 overexpression enhances the catalytic ability of ZDHHC6 [52].

Depalmitoylation

S-palmitoylation is typically dynamic and can be reversed by serine hydrolase superfamily members, mainly including APTs, α/β hydrolase domain-containing proteins (ABHDs) and PPTs (Fig. 2c-d) [53, 54]. APT is also named lysophospholipase (LYPLA), because they were initially discovered as lysophospholipases [55, 56]. Though the sequences and structures of APT1/2 are quite comparable, these two enzymes differ in localization and substrate specificity. APT1 predominantly accumulates on mitochondria while APT2 is mainly found on GA membrane [57]. APT1/2 play pivotal roles in many biological processes. For instance, signal transducer and activator of transcription-3 (STAT3) belongs to substrates of APT2, S-palmitoylation and depalmitoylation cycle of STAT3 changes its membrane localization and nuclear translocalization, thus contributing to immune cell differentiation [58].

The regulatory mechanisms of APTs are not yet fully characterized. APTs undergo PTMs including S-palmitoylation and ubiquitylation to achieve its localization and activity. It has been found that APT2 accumulation relies on its S-palmitoylation via ZDHHC3 and 7 on GA and ER respectively, palmitoylation-deficient APT2 would lose the catalytic ability [59]. Intriguingly, it seems that GA/cytosolic APTs pools depend on the rapid interconversion of S-palmitoylated and non-palmitoylated forms. Subsequent studies proposed that soluble APTs are active forms and these on GA are stable forms required to control the homeostatic status [60].

David Tse Shen Lin et al. reported the existence of additional thioesterases beyond APT1/2 by comparing half-lives of palmitate and protein, finding that APT1/2 knockdown did not block depalmitoylation of the PSD95. The result revealed that ABHD17 acted as a depalmitoylation enzyme for PSD95 and N-Ras [61]. Subsequent studies found that other ABHD family enzymes including ABHD8, 10, 16 A and 17 A/B/C all executed depalmitoylation activity [61–64]. Table 1 concludes these enzymes and their substrates. Indeed, ABHD17A/B/C all have strong depalmitoylating activity to PSD95. ABHD17 expressed in neurons selectively reduced PSD95 palmitoylation and synaptic clustering of PSD95 [65]. Elena Tortosa et al. found that ABHD17A/B/C catalyzed depalmitoylation of microtubule-associated protein-6 (MAP6), contributing to the shuttling of MAP6 between membranes and microtubules [66].

PPT1 is mainly located in lysosome, which serves as hydrolases to depalmitoylate proteins before their degradation [67]. PPT1 deficiency upregulates lysosomal pH value and damages enzyme activity, thus affecting the communication between lysosome and other organelles [68, 69]. PPT1 mutation causes infantile neuronal ceroid lipofuscinosis, due to undegraded lipofuscin accumulation in nerve cells [70]. PPT2 seems to be enzymatically inactive, it can hydrolyze unbranched molecule like palmitoyl-CoA, but cannot hydrolyze palmitoyl-cysteine or palmitoylated proteins [71].

As Table 1 shows, the number of depalmitoylating enzymes is much smaller than that of palmitoylating enzymes. Indeed, depalmitoylation enzymes belong to serine hydrolase superfamily, which includes more than 200 serine hydrolases identified so far, though not all the members carry depalmitoylation activity [53]. APT1/2 are expressed across nearly all tissues, allow hydrolyzing thioesters [72]. ZDHHCs recognize diverse protein substrates with distinct binding pockets, while depalmitoylases act as broad-spectrum “erasers”, recognizing thioester bonds rather than protein motifs. Additionally, membrane-embedded ZDHHCs require precise organelle localization, while soluble cytosolic APT1/2 can diffuse to multiple space.

Physiological effects of S-palmitoylation

Protein structure and conformation

S-palmitoylation exerts a crucial function in protein structure and conformation, contributing to alter the accessibility of active sites and disrupt interactions with other molecules (Fig. 3a). Epidermal growth factor receptor (EGFR) participates in receiving extracellular stimulation and initiating downstream signal cascades, C-terminal tail of EGFR is free from plasma membrane and extends into cytosol. Crucially, S-palmitoylation of EGFR C-terminal tail promotes its combination with plasm membrane by burying palmitate in lipid bilayer, impeding EGFR activation (Fig. 3a, left panel) [73]. Caspase-6 (CASP6) serves as a well-characterized example where S-palmitoylation induces direct conformational changes. CASP6 is found to be associated with cell apoptosis and the immune system, with four loops and the central β-sheet forming the activation site [74, 75]. There are two sites can be S-palmitoylated in CASP6: Cys277 close to the start of the β-sheet and Cys264 on loop 4. PA binding to Cys277 occupies active site, thus contributes to the blockage of activation and dimerization of CASP6. S-palmitoylation at Cys264 also blocks the active site of CASP6, relying on improving the flexibility of loop 4 and optimizing its dynamic interplay with loop 2 (Fig. 3a, middle panel) [76]. The Na⁺/Ca²⁺ exchanger-1 (NCX1) is engaged in cellular Ca²⁺ efflux in cardiac contractile cycle. XIP, a polybasic region of NCX’s regulatory loop, is usually sequestered by anionic phospholipid PIP2 and is responsible for NCX1 inactivation. Structural rearrangement of NCX1, facilitated by S-palmitoylation, leads to PIP2 hydrolysis, resulting in the release of XIP and promoting NCX1 inactivation (Fig. 3a, right panel) [77–79]. Collectively, these examples show that S-palmitoylation serves as a universal structural switch across functionally diverse proteins, highlighting its therapeutic potential for pathologies driven by protein mislocalization or dysregulated activation.

Fig. 3.

Physiological effects of S-palmitoylation. S-palmitoylation of proteins exerts crucial physiological effects. (a) S-palmitoylation regulates protein structure and conformation, contributing to alter the accessibility of active sites and disrupt interactions with other molecules. S-palmitoylation of EGFR C-terminal tail promotes its combination with plasma membrane. S-palmitoylation at Cys264 and Cys277 residues blocks the activation site of Caspase-6. S-palmitoylation of NCX1 modifies structural arrangement of NCX1 dimer. (b) S-palmitoylation has an important influence on proteins’ affinity to membranes and ensures them to shuttle among intracellular compartments. The initial assembling and trafficking site for AMPAR isomers is ER, S-palmitoylation leads to their accumulation and maturation in Golgi apparatus, reduces their stability to plasma membrane, and decreases their trafficking to lysosomes and degradation. (c) S-palmitoylation is capable of regulating cell metabolism through modulating substrates, transporters and enzymes. A variety of metabolic enzymes are observed to be S-palmitoylated, including HMG-CoA synthase and MDH2. CD36 is depalmitoylated by APT1 to initiate endocytic uptake of FAs, and FAs attaching to CD36 inactivates ZDHHC5. (d) S-palmitoylation affects cellular immune, especially in immune receptors and signaling proteins. S-palmitoylation of CD81 prolongs its residency and amplifies BCR-mediated signal to larger domains. PD-L1 is palmitoylated at Cys272 site, which enhances its stability and impairs anti-tumor effects of T cells. S-palmitoylation of STING in Golgi apparatus is essential for its assembly into multimeric complexes and the recruitment of downstream signaling factors. Abbreviations: AMPAR, α-amino-3-hydroxy-5-methyl-4-isox-azoleproprionic acid receptors; APTs, acyl-protein thioesterases; BCR, B cell receptor; EGFR, epidermal growth factor receptor; MDH2, malate dehydrogenase 2; NCX1, Na⁺/Ca²⁺ exchanger-1; PA, palmitic acid; PD-1, programmed cell death-1; PD-L1, programmed cell death ligand-1; STING, stimulator of interferon genes; TBK, TANK binding kinase; TCA cycle, tricarboxylic acid cycle

Protein trafficking and localization

S-palmitoylation critically influences protein membrane affinity and enables their shuttling between intracellular compartments. The hydrophobic palmitate bound to protein serves as an anchor, enhancing lipophilicity and facilitating interactions with membrane [20]. A typical example is its occurrence on synaptic proteins, where it is observed in over 40% of cases and emerges as a pivotal mechanism regulating synaptic transmission [80–82]. α-amino-3-hydroxy-5-methyl-4-isox-azoleproprionic acid receptors (AMPARs) contain 4 subunits (GluA1-4), distinct modifications enable their trafficking routes and synaptic functions. ER serves as the initial site for AMPAR subunit assembly and trafficking. All GluA1-4 subunits can be S-palmitoylated at two conserved cysteine residues, inhibiting lysosomal targeting and degradation (Fig. 3b) [83, 84]. GluA1/2 traffic from ER to GA, where AMPARs undergo S-palmitoylation and get accumulated and matured [82]. In addition, AMPARs depalmitoylation serves as a signal for their release from GA to plasm membrane [82]. Other studies have also revealed the relationship between protein S-palmitoylation and intricate trafficking and localization network, including N-methyl-D-aspartate receptors (NMDARs), γ-aminobutyric acid sub-type A (GABAA) receptors [85–87]. Deciphering the spatiotemporal control of palmitoylation cycles in receptor trafficking provides a fundamental blueprint for developing precision neuromodulators for diseases.

Metabolic regulation by S-palmitoylation

S-palmitoylation is capable of regulating cell metabolism through modulating substrates, transporters and enzymes. S-palmitoylation adds a hydrophobic palmitate tail to target proteins, increasing their affinity for membranes. A variety of metabolic enzymes are observed to be S-palmitoylated, including HMG-CoA synthase, malate dehydrogenase-2 (MDH2), as well as carbamoyl-phosphate synthetase-1 (CPS1) [88–90]. HMG-CoA synthase has two palmitoylation sites containing Cys166 and Cys305. S-palmitoylation of HMG-CoA synthase is required for its interaction with transcription factor peroxisome proliferator-activated receptor alpha (PPARα), and plays a key role in supporting mitochondrial metabolism [88, 91]. MDH2 is one of the key enzymes in tricarboxylic acid (TCA) cycle, S-palmitoylation on Cys138 residue of MDH2 increases its activity and enhances mitochondrial respiration (Fig. 3c) [89]. FAs transportation depends on CD36 S-palmitoylation and internalization. FAs binding to CD36 activates its downstream kinases, leading to ZDHHC5 phosphorylation and inactivation. CD36 subsequently undergoes APT1-mediated depalmitoylation to initiate endocytic uptake of FAs [92]. Blocking the dynamic S-palmitoylation and depalmitoylation cycle of CD36 eliminates FAs uptake, indicating its crucial role in cell metabolism [92]. Targeting S-palmitoylation in these metabolic transporters or enzymes may further correct energy imbalances in diseases.

S-palmitoylation in immune signaling

Published studies have elucidated the effects of S-palmitoylation in cellular immunity, especially in immune receptors and signaling proteins. Given its role in modulating protein stability and degradation, S-palmitoylation critically governs the function of immune receptors and checkpoints. The CD19/21/81 coreceptor complex enhances B cell receptor (BCR) signal transduction by prolonging it relation to lipid rafts. Antigen ligation to CD19/21/81 induces rapid S-palmitoylation of CD81, which prolongs its residency and amplifies BCR-mediated signal to larger domains [93]. Impairment of CD81 S-palmitoylation also disrupts its associated protein network, including immunoglobulin superfamily member-8 (IGSF8) and CD9 [94]. Programmed cell death ligand-1 (PD-L1) undergoes S-palmitoylation at Cys272, which stabilizes the protein [95]. Silencing of ZDHHC3 or blocking S-palmitoylation with 2-bromopalmitate (2-BP) promotes PD-L1 ubiquitination and lysosomal degradation, thus reducing intracellular PD-L1 levels and enhancing anti-tumor effects of T cells [95].

Nucleotide-binding oligomerization domain leucine-rich repeat and pyrin-domain-containing-3 (NLRP3) is a crucial component of inflammasome, which provides a critical target in inflammatory diseases. S-palmitoylation of NLRP3 is catalyzed by ZDHHC12, promoting its degradation via chaperone-mediated autophagy pathway. ZDHHC12 deficiency is found to enhance inflammatory symptoms and even cause endotoxic shock in mice [96]. In addition, S-palmitoylation of stimulator of interferon genes (STING) protein in GA is essential for its multimerization and recruitment of downstream signaling factors. Pharmacological inhibition of STING S-palmitoylation could attenuate STING-mediated inflammatory cytokine production and immune response [97]. Collectively, targeting S-palmitoylation of immune response represents a promising strategy for the treatment of autoinflammatory disease or strengthen immunotherapies of tumors.

Effects of S-palmitoylation in human diseases

Organized and reversible S-palmitoylation achieve remarkable roles in coordination of protein functions and the precise orchestration of biological processes. Increasing studies have focused on how dysregulated S-palmitoylation afflicts pathological consequences and how to target S-palmitoylation as candidate therapeutic strategies. Here we systematically summarize the scene of S-palmitoylation in human diseases (Fig. 4).

Fig. 4.

Effects of S-palmitoylation in human diseases. S-palmitoylation has been implicated in the pathogenesis of diverse diseases including cancers, cardiovascular diseases, metabolic diseases, immunological diseases, nervous system diseases and mental diseases. Abbreviations: AKAP150, A-kinase anchoring protein-150; AMI, acute myocardial infarction; AML, acute myeloid leukemia; APTs, acyl-protein thioesterases; ATGL, adipose triglyceride lipase; BACE1, β-site APP cleaving enzyme-1; BC, breast cancer; BCa, bladder cancer; CASP6, caspase-6; CLL, chronic lymphocytic leukemia; DLBCL, diffuse large B-cell lymphoma; ERα, estrogen receptor-α; GBM, glioblastoma; GC, gastric carcinoma; GLUT4, glucose transporter 4; GSDMD, gasdermin D; HCC, hepatocellular carcinoma; HD, Huntington disease; HTT, huntingtin; IBD, inflammatory bowel disease; IFN-α, interferon-α; IRHOM2, inactive rhomboid protein-2; NLRP3, nucleotide-binding oligomerization domain leucine-rich repeat and pyrin-domain-containing-3; NOD2, nucleotide oligomerization domain-like receptors 2; NSCLC, non-small cell lung cancer; OC, ovarian cancer; PA, palmitic acid; PATs, protein acyltransferases; PD, Parkinson’s disease; PDAC, pancreatic ductal adenocarcinoma; PDE10A, phosphodiesterase-10 A; PPT1, palmitoyl-protein thioesterase 1; PRDX6, peroxiredoxin-6; SNAP25, synaptosome associated protein-25; STAT3, signal transducer and activator of transcription 3; STING, stimulator of interferon genes; SYT1, synaptotagmin-1; TGR5, Takeda G protein-coupled receptor-5; 5-HT1AR, serotonin 1 A receptor

Cancers

Glioblastoma (GBM)

GBM is a highly malignant and heterogeneous tumor, with a median survival of approximately one year. The role of S-palmitoylation and its metabolic implications in GBM are actively investigated. For example, S-palmitoylated variant of solute carrier family 1 member-3 (SLC1A3) is downregulated in GBM, significantly impairing glutamate uptake [12]. GLUT1 requires ZDHHC9-mediated S-palmitoylation for membrane retention and glucose transport function. Disruption of GLUT1 S-palmitoylation inhibits glycolysis, blocks cell proliferation and suppresses GBM tumorigenesis [98]. Additionally, ZDHHC19-mediated S-palmitoylation of SMAD family member-3 (Smad3) is confirmed to activate transforming growth factor (TGF) signaling pathway in GBM. The interaction between Smad3 and E1A binding protein P300 (EP300) promotes mesenchymal-like transition of GBM [99]. ZDHHC8 mediates S-palmitoylation of the cystine/glutamate antiporter SLC7A11 at Cys327 in GBM, contributing to maintain its stability via reducing ubiquitination level [41]. This mechanism is essential for ferroptosis resistance in GBM tumorigenesis, suggesting a potential therapeutic method for GBM.

Breast cancer (BC)

BC remains the leading cause of cancer-related mortality in women worldwide and exhibits strong reliance on FAs metabolism. S-palmitoylation directly modulates energy metabolism in BC cells by regulating lipid metabolic pathways. Claudin-6 (CLDN6), a regulator of FA biosynthesis, was recently found to downregulate sterol regulatory element-binding protein-1 (SREBP1) transcription, subsequently reducing de novo palmitic acid synthesis. This metabolic alteration hinders Ras S-palmitoylation and suppresses BC progression [13]. Plasm membrane and mitochondrion EGFR (pm/mtEGFR) overexpression in BC activates FASN, elevating de novo palmitate production. Subsequent mtEGFR palmitoylation promotes tyrosine kinase inhibitor (TKI) resistance [100]. Besides, S-palmitoylation has an impact on survival, proliferation and apoptosis of BC cells. Neurotensin receptor-1 (NTSR1) has two S-palmitoylation sites: Cys381 and Cys383 residues. Mutating these sites in BC cells decreases NTSR1 membrane localization, reduces its interaction with Gαq/11, and rescues BC cells from apoptosis [101]. ZDHHC3 mediates integrin beta 4 (ITGβ4) S-palmitoylation, which maintains its localization within lipid raft microdomains and promotes BC cell invasiveness [102]. S-palmitoylation is also anticipated to influence tumor immunity in BC. ZDHHC9 catalyzes S-palmitoylation of PD-L1 to enhance its stability [103, 104]. Disrupting PD-L1 S-palmitoylation, either through inhibiting ZDHHC9 or inducing specific mutations, could sensitize BC cells to T cell killing effect, providing a potential strategy to improve anti-PD-L1 therapeutic efficacy [103].

Non-small cell lung cancer (NSCLC)

NSCLC ranks among the most prevalent malignancies. Recent research reveals the tight association between EGFR S-palmitoylation and NSCLC. In lung adenocarcinoma, blocking EGFR S-palmitoylation inhibits phosphatidylinositol 3 (PI3K) kinase activation, reduces MYC expression levels and decreases cell proliferation, ultimately attenuating tumor growth [105]. Azhar Ali et al. found that mutated EGFR signaling in TKI-resistant tumor cells depends on EGFR palmitoylation and can be targeted by Orlistat. Inhibition of FASN with Orlistat induces ubiquitination of EGFR, and abrogates its mutant signaling [106]. Furthermore, genomics and proteomic database analyses reveal that ZDHHC5-catalyzed S-palmitoylation of INCENP (a core component of the chromosomal passenger complex) modulates cancer stem cells (CSCs) in lung adenocarcinoma [107]. Collectively, these findings underscore the critical role of S-palmitoylation in NSCLC pathogenesis, where targeting EGFR palmitoylation or modulating INCENP palmitoylation disrupts oncogenic signaling and tumor growth.

Digestive system tumors

Gastric carcinoma (GC) usually exhibits rapid progression and poor prognosis. Nuclear factor erythroid 2-related factor-2 (Nrf2) serves as the main regulator of cellular homeostasis during oxidative stress, and its dysregulation contributes to GC carcinogenesis and progression [108]. Liu et al. found that high ZDHHC2 level promoted S-palmitoylated Nrf2 abundance in GC, and this modification blocked ubiquitin-proteasome-related degradation of Nrf2, consequently leading to GC growth [109]. These results extend candidates for possible pharmacological targets for GC treatment.

TGF-β is found to stimulate S-palmitoylation of hexokinase-1 (HK1), which facilitates its secretion via large extracellular vesicles. Hepatocellular carcinoma (HCC) cells can capture these vesicle HK1, contributing to glycolysis acceleration and HCC progression [110]. Currently, Sorafenib is the first-line therapeutic regimen for advanced HCC, but the resistance remains a rigorous challenge. Sun et al. revealed that aberrant upregulation of proprotein convertase subtilisin/kexin-9 (PCSK9) induced AKT-S473 phosphorylation and promoted sorafenib resistance in HCC, suggesting a promising target for HCC treatment [111].

According to palmitoylation proteomics studies, pancreatic ductal adenocarcinoma (PDAC) is found to be rich with proteins that have known S-palmitoylation targets, suggesting aberrant S-palmitoylation may impact pancreatic tumorigenesis [112]. Both cytoskeleton-associated protein-4 (CKAP4) and LDL receptor-related protein-6 (LRP6) could be S-palmitoylated by ZDHHC2, which induces them to localize in detergent-resistant membrane fractions and activates PI3K-AKT pathway, thus promoting cell proliferation in PDAC [113]. One remarkable study reveals that ZDHHC20 is abnormally upregulated in PDAC, relating to poor prognosis. YTH domain-containing family protein-3 (YTHDF3), one of the main m6A readers, could be S-palmitoylated by ZDHHC20, inhibiting its chaperone-mediated autophagic degradation and causing accumulation of oncogenic product MYC [114].

In colorectal cancer (CRC), interferon gamma receptor-1 (IFNGR1) is S-palmitoylated on Cys122 residue, participating in IFN-γ and MHC-I signaling integrity. Optineurin protects IFNGR1 from lysosomal sorting and degradation, which impairs IFN-γ and MHC-I signal pathways and drives immune evasion and intrinsic immunotherapy resistance in CRC [115]. Zhang et al. recently reveals that acyl-CoA oxidase-1 (ACOX1) is significantly downregulated in CRC, correlating to inferior prognosis in CRC patients. ACOX1 downregulation increases the S-palmitoylation of β-catenin, facilitating its stabilization and subsequent signal pathway, thus promoting CRC progression [116].

Urinary system tumors

The incidence of renal cell carcinoma (RCC) is increasing annually. STING, an innate immune sensor, plays an essential role in maintaining mitochondrial reactive oxygen species (ROS)/calcium homeostasis. Critically, its interaction with downstream voltage dependent anion channel-2 (VDAC2) depends on S-palmitoylation. In RCC, using ZDHHC inhibitors such as 2-BP to block STING binding to VDAC2 efficiently suppressed tumor cell growth [117].

Separately, emerging evidence implicates the oncogenic role of ZDHHC9 and it mediated Bip S-palmitoylation in bladder cancer (BCa). Bip S-palmitoylation enhances its ER retention, reduces protein degradation, and consequently maintains cellular homeostasis. This study also confirms that ZDHHC9 promotes drug resistance in BCa, highlighting its potential as a novel therapeutic target [118].

Genital system tumors

S-palmitoylation has been proposed to modulate cell survival and proliferation in ovarian cancer (OC). ZDHHC18-mediated S-palmitoylated of MDH2 is required for mitochondrial respiration and OC cell proliferation [89]. ZDHHC12-mediated claudin-3 (CLDN3) S-palmitoylation promotes OC progression. Conversely, impaired S-palmitoylation of CLDN3 disrupts its membrane anchoring and stability, ultimately exerting inhibitory effects on proliferation [119]. ZDHHC12 further regulates mitochondrial function and ROS homeostasis, knockdown via siRNA or inhibition using 2-BP/FASN inhibitors enhances cisplatin’s anti-tumor efficacy in OC [120].

Multiple S-palmitoylated proteins are engaged in regulating cell proliferation in prostate cancer (PCa). Src kinase requires N-terminal palmitoylation for oncogenic transformation through differential localization [121]. While androgen receptor (AR) promotes proliferation and evades apoptosis upon hormonal ligand binding, palmitoylation-dependent plasm membrane localization of AR8 reduces PCa cell proliferation and induces apoptosis [122]. Additionally, ZDHHC7 is found to inhibit AR gene transcription and consequently abolishes AR signaling in PCa cells [123]. Collectively, S-Palmitoylation bidirectionally regulates malignancy in genital tumors by modifying key proteins. These findings underscore pathological significance of the palmitoylation network, revealing ZDHHCs as promising therapeutic targets.

Hematological tumors

Emerging research has extensively investigated the functional significance of S-palmitoylation in the pathogenesis and progression of hematological malignancies. In acute myeloid leukemia (AML), oncogenic NRas requires S-palmitoylation for leukemogenesis [14]. The nuclear translocation of phospholipid scramblase-1 (PLSCR1), a multipalmitoylated protein, is identified to be necessary for cell differentiation in AML. The S-palmitoylation/depalmitoylation status of PLSCR1 act as a switch to regulate its destination either to plasm membrane or nucleus [124]. Clinically relevant to poor prognosis, internal tandem duplication within FLT3 (FLT3-ITD) undergoes ZDHHC6-mediated palmitoylation. Critically, blocking this process redirects FLT3-ITD to plasm membrane, rewiring AKT signaling to accelerate leukemia progression [125]. Palmitoylated proteins on AML-derived extracellular vesicle surface activates Toll-like receptor 2, driving Akt/mTOR-dependent induction of myeloid-derived suppressor cells (MDSC) [126]. Beyond signaling molecules, CD36-mediated palmitate uptake enhances MYD88 palmitoylation, thereby activating NF-κB and suppressing anti-tumor effects of T cells [127]. Shao et al. demonstrated that ZDHHC21 regulates OXPHOS hyperactivity in AML cells. Inhibiting ZDHHC21 suppresses OXPHOS, markedly eradicating AML blasts [128].

Recent studies also revealed the critical role of S-palmitoylation in other types of leukemia. ZDHHC3-mediated palmitoylation of promyelocytic leukemia/retinoic acid receptor alpha (PML/RARα) regulates its oncogenic transcriptional activity. ZDHHC3 depletion or inhibition significantly arrests the malignant progression of acute promyelocytic leukemia (APL), especially drug-resistant APL [129]. ZDHHC14 is found to be overexpressed in acute biphenotypic leukemia and subsets of AML, which inhibits cellular differentiation and is related to inferior prognosis [130].

In terms of lymphoid malignancies, membrane proteins are less palmitoylated in chronic lymphocytic leukemia (CLL) cells. APT1/2-CD95 interaction is upregulated, thereby promoting depalmitoylation and impairing CD95-mediated apoptosis in CLL [131]. G protein subunit alpha-13 (GNA13) palmitoylation maintains its stability and plasm membrane localization in germinal center B-cell diffuse large B-cell lymphoma (GCB-DLBCL). Through this modification, GNA13 downregulates B-cell lymphoma-2 (BCL2), potentially sensitizing tumors to BCL2 inhibitors [132]. Additionally, ZDHHC21 acts as a tumor suppressor by palmitoylating FASN at Cys1317, directly impairing FASN stability and FA synthesis to inhibit DLBCL cell growth [133]. In Burkitt lymphoma (BL), MYC induces ZDHHC11/11B to maintain high MYB levels, contributing to the oncogenic MYC-miR-150-MYB axis in BL [134].

Other tumors

S-palmitoylation of GNAQ and GNA11 (GNAQ/11) has already been discovered, but its implication in uveal melanoma (UM) remains indistinct. Zhang et al. reveals that the loss of GNAQ/11 S-palmitoylation in UM cells obviously inhibits cell survival and proliferation. Using S-palmitoylation inhibitors could suppress GNAQ/11Q209L-induced malignant transformation in UM [135].

In malignant mesothelioma, some selective inhibitors are described to block transcriptional enhanced associate domain (TEAD) auto-palmitoylation and disrupt interaction between YAP/TAZ and TEAD, serving an important role in cell survival and proliferation [136]. This may provide a novel way targeting Hippo-YAP pathway in malignant mesothelioma.

Cardiovascular diseases

Acute myocardial infarction (AMI)

AMI carries heavy economic burden for society due to its high mortality and disability rates. After coronary blood flow is blocked, cardiomyocytes suffer various death modes including pyroptosis. Based on AMI mice model, GSDMD S-palmitoylation is found to promote its cytomembrane localization, resulting in IL-18 release and myocardial pyroptosis [137]. In addition, short-term high PA diet aggravates myocardial infarction, and treating with disulfiram would relieve this process [137]. The S-palmitoylation of STING, a downstream signaling effector of cGAMP, plays an essential role in damage-associated molecular patterns in acute immune response after cell death. STING inhibitors bind to STING and block its S-palmitoylation, thus preventing STING multimer assembly for TANK binding kinase-1 (TBK1) phosphorylation [138].

Myocardiopathy

Recent research proposes that S-palmitoylation is also implicated in diabetic cardiomyopathy. In a high-fat diet mice model, Takeda G protein-coupled receptor-5 (TGR5) deletion promotes CD36 localization to plasm membrane and upregulates its S-palmitoylation level, contributing to lipid accumulation and cardiac dysfunction [139]. NLRP3 activation relies on the removal of its palmitoyl groups, which exerts a vital effect in septic cardiomyopathy (SCM). Exposure of lipopolysaccharide is found to cause oxidative stress, inflammation, mitochondrial disorder, as well as cell apoptosis in cardiomyocytes, dealing with Vaccarin would alleviate the injury via inducing NLRP3 S-palmitoylation [140].

Metabolic diseases

Fatty liver

Inactive rhomboid protein-2 (IRHOM2) exerts an important function in nonalcoholic steatohepatitis (NASH) via regulating inflammation, immunoreaction, as well as cell survival. IRHOM2 is S-palmitoylated at Cys476 by ZDHHC3, facilitating its stabilization and cytomembrane translocation. Moreover, metabolic stress upregulates ZDHHC3, contributing to S-palmitoylated IRHOM2 accumulation by blocking its ubiquitination and ubiquitin-proteasome related degradation. This mechanism markedly accelerates the development of NASH [141]. In metabolic dysfunction-associated steatohepatitis, activating transcription factor-3 (Atf3) could attenuate hepatocyte steatosis by regulating macrophages’ glucose-FA cycle. Atf3 is found to increase CD36 S-palmitoylation and subsequently promote FAs uptake and oxidation [142].

Diabetes

Diabetes is a major chronic disease worldwide, severely afflicts life quality of patients. Abnormal lipolysis dysregulation may contribute to β-cell dysfunction in type 2 diabetes islets. Adipose triglyceride lipase is the principal lipase of human β-cells, its S-palmitoylation protects the downstream factor SNARE protein syntaxin-1a from proteasomal degradation and supports insulin secretion [143]. Skeletal muscle insulin resistance is one of a major contributor to type 2 diabetes, due to that GLUT4 transports to plasm membrane and facilitates glucose uptake into muscle. Mitigating ER stress or inhibiting protein S-palmitoylation rescues GLUT4 perinuclear sorting, providing potential therapies directly targeting GLUT4 translocation [144]. Diabetic neuropathy causes severe pain and impaired motility, which is identified with S-palmitoylation. S-palmitoylation of peroxiredoxin-6 regulates the activity of AE3 anion exchanger enzyme, leading to its protective effect against diabetic neuropathy [145].

Immunological diseases

Inflammatory bowel disease (IBD)

High fat diet is commonly recognized as an important triggering factor for IBD. PA enters intestinal epithelial cells through CD36 transportation and participates in STAT3 S-palmitoylation cycle (STAT3 S-palmitoylation catalyzed by ZDHHC7 anchors it to cell membrane, and its depalmitoylation catalyzed by APT2 facilitates nuclear translocation of it) [146]. Wei et al. demonstrates that PA upregulates STAT3 S-palmitoylation cycle, thus enhancing Th17 differentiation and exacerbating colitis development. Using 2-BP, APT2 inhibitor ML349, or knocking ZDHHC7 could disrupt STAT3 S-palmitoylation cycle and block PA-induced inflammatory in colitis [147]. Nucleotide oligomerization domain-like receptors 2 (NOD2) S-palmitoylation based on zDHHC5 is revealed to facilitate its membrane recruitment to promote proinflammatory effect, and it is depalmitoylated by ABHD17. In epithelial cells, inhibiting ABHD17 increases plasm membrane localization of NOD2, leading to NF-κB activation and pro-inflammatory cytokines production, consequently promoting Crohn’s disease [148].

Systemic lupus erythematosus (SLE)

Toll-like receptor 9 (TLR9) plays intricate roles in initiating innate immune response, and is correlated to autoimmune diseases including systemic sclerosis, psoriasis, as well as SLE [149]. Depalmitoylation of TLR9 is catalyzed by PPT1 in lysosome, which regulates cytokine response of macrophages, B cells and dendritic cells. Additionally, PPT1 inhibitor HDSF is found to suppress IFN-α in SLE [150].

Infection diseases

NOD1/2 are intracellular pattern-recognition proteins for antimicrobial defense. S-palmitoylation of NOD1/2 by ZDHHC5 is required for recognizing peptidoglycans, recruiting bacteria-containing endosomes and promoting effective immune response [151]. This S-palmitoylation mechanism of pattern recognition receptors have been observed in several cells, including macrophages and monocytes [151]. Pathogens likewise exploit S-palmitoylation to enhance infectivity and evade immune detection. Kiyoshi Kawasaki et al. revealed that S-palmitoylation of Salmonella lipid A, one core component of outer membrane lipopolysaccharide (LPS), attenuates TLR4 signaling and suppresses host pro-inflammatory responses [152, 153].

S-palmitoylation of viral proteins was first reported in 1970s [154], and recent pandemic of SARS-CoV-2 attracts increasing focus for this field. Viruses usually exploit host S-palmitoylation machinery due to their deficiency of PATs. Many viral proteins including spike glycoproteins and viroporins undergo S-palmitoylation, which is required for host-endosome membrane fusion [155]. SARS-CoV-2 virions have a lipid bilayer that is rich in spike glycoproteins for target-cell fusion. S-palmitoylation of spike glycoproteins by ZDHHC20 mainly occurs in ER and GA, allowing viruses formation with enhanced fusion capacity [156]. Enveloped viruses such as herpes simplex virus (HSV) and influenza virus, are rich in S-palmitoylated spikes glycoproteins, which contribute to host receptor engagement and endocytosis triggering [157]. Hyaluronic acid (HA) of influenza A virus is S-palmitoylated at two cysteine residues near to cytoplasmic tail, driving conformational changes and membrane fusion [157]. Additionally, the amphiphilic helix of matrix protein M2 undergoes S-palmitoylation, ensuring the proton channel activity of M2 and facilitating viral genome transport to the host nucleus [158].

Nervous system diseases

Huntington disease (HD)

Abnormal trafficking of mutant HTT has been identified as an early and consistent feature of HD. The S-palmitoylation and distribution of HTT are regulated by ZDHHC17 (also named huntingtin interacting protein 14 (HIP14)). Anat Yanai et al. revealed that decreased S-palmitoylation level of HTT accelerates inclusion formation, contributing to HD pathogenesis [159]. Additionally, ZDHHC17 shows remarkable substrate specificity for neuronal proteins, such as SNAP25, PSD95, synaptotagmin-1 (SYT1) and glutamic acid decarboxylase-65 (GAD65) [160]. ZDHHC17-deficient mice were found to exhibit neuropathological defects, including increased striatum cell death, striatum degeneration, and cortex degeneration [161]. ZDHHC17 also catalyzes S-palmitoylation of CASP6, which is found to block its substrate-binding groove and inhibit dimerization. This mechanism is crucial for CASP6 activation and helps to restore neuronal function in HD [76]. ZDHHC13 is a homolog of ZDHHC17 with similarly strong activity to promote HTT palmitoylation. The interaction of ZDHHC13 with mutant HTT was found to be dramatically diminished in HD [162]. ZDHHC13-deficient mice also show HD-like striatal pathology, with progressive reductions in cortical volume, striatal, as well as brain weight [25].

Excessive activation of glutamate receptors plays a vital role in the pathogenesis of HD. The NMDAR is upregulated in HD, triggering cell death and driving HD neurodegeneration [163]. NMDAR displays two distinct C-terminal clusters of S-palmitoylation. S-palmitoylation of the first cluster enhances tyrosine phosphorylation, increasing the surface stability of NMDAR. While S-palmitoylation of the second cluster reduces its surface expression [25]. Sequestosome 1 (SQSTM1)/p62, a receptor for autophagy, can be S-palmitoylated and direct to lysosome. Recent study found that SQSTM1/p62 palmitoylation is reduced in HD, which may be responsible for autophagy defect in HD [164]. Based on proteomic analysis of transporting vesicle in corticostriatal projecting neurons, S-palmitoylation is identified to restore defective huntingtin trafficking. ML348 could inhibit APT1 to upregulate S-palmitoylation level and restore axonal transport, thus reversing neuropathology, anxio-depressive behaviors, as well as locomotor deficits [165]. Therefore, targeting S-palmitoylation may provide new ideas and directions for the treatment and prevention of HD.

Alzheimer’s disease (AD)

AD afflicts an increasing number of older patients, with pathological features including synaptic dysfunction, neurofibrillary tangles and amyloid plaques, leading to cognitive impairment. Many studies have indicated the role of S-palmitoylation in AD. Regarding palmitoylating enzymes, for example, ZDHHC21 p.T209S (a rare gain-of-function variant) mutation enhances S-palmitoylation of β amyloid precursor protein in hippocampus, followed by increasing of Aβ40 and 42 [166]. ZDHHC21 also catalyzes S-palmitoylation of transient receptor potential vanilloid-2 at Cys277 residue, blocking this process could improve microglial Aβ phagocytosis [166]. ZDHHC7 expression and protein S-palmitoylation were increased in hippocampi of AD mice and AD patients. Using 2-BP could reduce the Aβ deposition and counteract cognitive deficits in mouse model [167]. SELENOK participates in microglial migration, phagocytosis and immune regulation, potentially associated with AD pathology. SELENOK is found to affect CD36 S-palmitoylation, which regulating CD36 to anchor to plasm membrane and consequently impacting Aβ phagocytosis in microglial cells. High Se level upregulates SELENOK expression and CD36 S-palmitoylation, thus promoting microglial Aβ phagocytosis and mitigating the progression of AD [168].

In terms of palmitoylation substrate, β-site APP cleaving enzyme-1 (BACE1) initiates Aβ production, contributing a critical role in AD pathogenesis. BACE1 has been found to be S-palmitoylated at four juxtamembrane cysteine residues. In AD mouse models, lack of BACE1 S-palmitoylation impaired synaptic activity-induced Aβ production, consequently reducing cerebral amyloid burden and mitigating cognitive deficits [169]. Xavier Meckler et al. found S-palmitoylation of two γ-secretase subunits, APH1aL and nicastrin, which enables cleavage of amyloid precursor protein to generate Aβ. S-palmitoylation deficiency of APH1aL and nicastrin significantly reduced insoluble Aβ [170]. Zhang et al. found increased S-palmitoylation of PSD95 in AD, cattle encephalon glycoside and ignotin treatment markedly inhibited PSD95 palmitoylation and enhanced expression of synaptic-associated proteins [171].

Parkinson’s disease (PD)

SYT11 and α-synuclein are involved in vesicle trafficking, and play crucial roles in PD. SYT11 S-palmitoylation is found to disrupt α-synuclein homeostasis in neuron and upregulate aggregation-prone monomeric of α-synuclein. The result indicates that S-palmitoylation-mediated increases of SYT11 might facilitate pathological aggregation of α-synuclein in PD [172]. Synaptic abnormalities of PD are delayed in females, mainly due to that estradiol improves α-synuclein homeostasis via local estrogen receptor-α (ERα) [173]. ERα S-palmitoylation is crucial for its distribution, and using depalmitoylation inhibitors could increase ERα’s availability and ameliorate synaptic plasticity in PD [174].

Mental diseases

Depressive disorder

S-palmitoylation has a close linkage to mental disease, the deep mechanisms are in focal learning currently. For example, 5-HT1AR is implicated in depressive disorder, which undergoes S-palmitoylation in human. ZDHHC21 deficiency reduces 5-HT1AR S-palmitoylation and impairs receptor signaling. Downregulation of 5-HT1AR S-palmitoylation plays an important role in depressive disorder, providing a delightful method targeting the restoration of 5-HT1AR S-palmitoylation for depression treatment [175]. Separately, A-kinase anchoring protein-150 (AKAP150) S-palmitoylation is upregulated in mice with induced depressive-like behaviors. Blocking AKAP150 and peptide Ht-31 interactions or inhibiting palmitoylation attenuates synaptic transmission and rescues depressive phenotypes [176]. Notably, chronic pain-induced depression also correlates with S-palmitoylation. Peripheral spared nerve injury activates astrocyte to release IL-6, which facilitates S-palmitoylation of PSD95. This modification promotes GluR1/NR2B recruitment to synapses, further exacerbating depression-like behaviors [177].

Anxiety disorder

In amygdala, endocannabinoid (eCB) function relies on its tyrosine phosphorylation, and eCB signaling deficiency contributes to stress-induced anxiety. LMO4 is an endogenous inhibitor of protein tyrosine phosphatase 1B (PTP1B), decreased LMO4 S-palmitoylation could reduce eCB level through elevating PTP1B activity, which contributes to rescue anxiety behaviors [178]. S-palmitoylation deficiency of gephyrin is observed in highly anxious rats, accompanied with ZDHHC12 dysfunction. Anxiolytic drug diazepam could increase gephyrin S-palmitoylation and activate ZDHHC12 [179, 180]. Collectively, protein S-palmitoylation shows critical roles in orchestrating anxiety behaviors, which might serve potential strategies for anxiolytic intervention.

Schiziaophren

Phosphodiesterase-10 A (PDE10A) acting as a potential therapeutic target has gained considerable attention currently. PDE10A is a membrane-bound phosphodiesterase and is transported based on medium spiny neuron axons and dendrites, which relies on its S-palmitoylation by ZDHHC7 and 19 [181]. Approximately 30% 22q11.2 deletion carriers develop schizophrenia, with a disrupted ZDHHC8 gene. ZDHHC8 catalyzes S-palmitoylation of proteins that are involved in axonal growth and branching, and impaired terminal arborization causes a reduction in synaptic connections, consequently affecting functional connectivity and working memory [182].

Inhibitors targeting S-palmitoylation and depalmitoylation

These days, accumulating studies have shown that knockout or knockdown of individual ZDHHCs downregulates S-palmitoylation and impairs associated biological processes [58, 73]. Inhibitors targeting S-palmitoylation represent a promising class of compounds worked by either blocking the enzymatic activity of PATs or disrupting the S-palmitoylation of specific target proteins. Though the complicated intrafamilial regulatory mechanism and functional redundancy among ZDHHC-PATs hinder the study of highly selective inhibitors, chemical inhibitors allow assessment of palmitoylation-targeted therapeutic strategies and have gained booming focus. Currently, S-palmitoylation inhibitors are mainly divided into two classes: ZDHHCs inhibitors and non-enzymatic proteins. The inhibitors of ZDHHCs are further sorted into lipid-like and non-lipid inhibitors [9]. Nevertheless, the development of these inhibitory agents remains at a preliminary stage, detailed pharmacological effects and potential clinical application still need to be evaluated. Here we describe two non-selective inhibitors that have been widely studied and one FAD approved drug, other current inhibitors of S-palmitoylation and depalmitoylation are summarized in Table 2.

Table 2.

Inhibitors targeting S-palmitoylation and depalmitoylation

Inhibitor	Targets	IC50	Mechanisms
2-BP	ZDHHCs, APTs	10–15 µM	2-BP is considered as a palmitic analogue, which covalently binds to active-site cysteine of ZDHHCs and inhibits enzymatic activity. It is non-selective and has been found to perturb almost all ZDHHCs.
CMA	ZDHHCs	1.35 µM (ZDHHC20)	Using an acrylamide warhead to replace α-halo fatty acid of 2-BP could reduce toxicity and avoid inhibition of the S-palmitoylation eraser enzymes including APT1/2.
MY-D-2	ZDHHC7	38 µM (ZDHHC7)	Palmitic analogue
MY-D-4	ZDHHC3, 7, 20	4.4 µM (ZDHHC7), 50 µM (ZDHHC3)	It is an almitic analogue and its cysteamine moiety mimicking acyl-CoA.
Compound 8i	ZDHHCs	50 µM	A covalent S-palmitoylation inhibitor using 2-BP and CMA as the lead compounds.
Compound IV	ZDHHCs	0.5–11.8 µM	It is a non-lipid and reversible inhibitor, its specific binding site within the ZDHHC protein remains unidentified.
Artemisinin	ZDHHC6	27.9 µM	It covalently inhibits ZDHHC6 as a nonlipid palmitoylation inhibitor, thereby reducing S-palmitoylation of the oncogenic protein NRAS.
Curcumin	ZDHHC3		Unsaturated α, β-carbonyl groups of curcumin facilitate binding to the nucleophilic sulfhydryl group of cysteine residues.
Bis-piperazine backbone compounds 13 and 25	ZDHHC9, Erf2 [1]	50–150 µM	It inhibits Erf2 autopalmitoylation.
TTZ-1	ZDHHC2, 15, 17	15 µM (ZDHHC2)	It effectively inhibits the self-palmitoylation of ZDHHC2 as well as the S-palmitoylation of its substrates SNAP25 and PSD95.
TTZ-2	ZDHHC2, 15, 17	19 µM (ZDHHC2)	It effectively inhibits the self-palmitoylation of ZDHHC2 as well as the S-palmitoylation of its substrates SNAP25 and PSD95.
Disulfiram	ZDHHCs		Disulfiram can inhibit S-palmitoylation of NLRP3 at Cys126, which is important for localization and activation of NLRP3.
Palmostatin B	APT1, 2, ABHD16, 17	5.4 nM (APT1), 19.58 nM (APT2), ABHD16 (100 nM)	It possesses a β-lactone core and is the most potent compound in inhibiting APT1.
Palmostatin M	APT1, 2, ABHD17	2.5 nM (APT1), 36.74 nM (APT2)	It perturbs the signaling of Hand N-Ras by inhibiting depalmitoylation, which in turn led to decrease of MAPK signaling and partial phenotypic reversion.
HDFP	APT1, 2, PPT1, ABHD10, and ABHD17 A/B/C		A versatile serine hydrolase inhibitor.
ML211	APT1, 2	17 nM (APT1), 30 nM (APT2)	A triazoluria inhibitor.
ML348	APT1	280 nM	In the eutectic structures of APT1 and ML348, the aniline carbonyl group made two water molecules mediated hydrogen bonds.
ML349	APT2	120 nM	A crystal structure of human APT2 bound.
ABL117	ABHD10, 16 A, 17	210 nM	Targeting the uncharacterized serine hydrolase ABHD10.
ABL303	ABHD10	30 nM	It is developed by introducing isopropyl groups on the carbamates and moving the C-3 methyl group to the para position of the benzene ring.
MIDA-boronates	ABHD10	10 µM	It exhibits complete inhibition and demonstrated a near-total inactivation of ABHD10.
12-thiazole abietanes	ABHD16	3.4 µM	It belongs to the abietane family, and is a reversible inhibitor of ABHD16A.
DQ661	PPT1		It is a novel dimeric quinacrine that affects multiple lysosomal functions and mTORC1 activity by specifically targeting PPT1.
DC661	PPT1		Comparing to DQ661, it can maintain its inhibitory activity against PPT1 in acidic media.
Ezurpimtrostat	PPT1	0.22 µM	It triggers the accumulation of free Zn2+ ions within lysosomes and impairs tissue protease activity by inhibiting the PPT1.

^[1] Erf2 is the Saccharomyces cerevisiae PAT responsible for catalyzing the palmitoylation of Ras2

2-BP

2-BP is considered as a palmitic analogue, exerting its inhibitory effect through covalent modifying active-site cysteine of ZDHHCs and inhibiting enzymatic activity [183]. It is a “pan-inhibitor”, and has been found to perturb almost all ZDHHCs [7]. 2-BP is converted to 2-BP-CoA in cytoplasm, and then binds to ZDHHCs. Though the efficacy of 2-BP is less than free palmitate, its reactivity is dramatically strengthened once conjugating to CoA [184]. 2-BP is widely used to disturb S-palmitoylation of targeted proteins to examine their biological roles. For instance, Wei et al. applied 2-BP to reduce S-palmitoylation of autophagy related (ATG) proteins, demonstrating that ATG16L1was S-palmitoylated on Cys153 residue to promote LC3-II production and autophagosome formation [185].

In preclinical experiment, 2-BP has shown therapeutic potential in multiple diseases. For example, 2-BP inhibited PD-L1 palmitoylation activates anti-tumor immunity in CRC mice [95]. Targeting β-catenin palmitoylation via 2-BP inhibits β-catenin-dependent tumor growth and reduces the viability of CRC cells [116]. However, 2-BP has severe drawbacks to limit its clinical applications: promiscuity and cytotoxicity. Apart from various ZDHHCs, 2-BP and 2-BP-CoA are also found to impact enzymes related to lipid and glucose metabolism, such as FA CoA ligase, glucose-6-phosphatase and NADPH cytochrome-c reductase [186]. Besides, 2-BP interferes with depalmitoylation enzymes APT1/2. Laurence Abrami et al. found that membrane binding of APT2 protected it from proteasomal degradation, its interaction with membrane relied on S-palmitoylation via ZDHHC3/7. This mechanism suggests that 2-BP could impair the stabilization and function of APT2 [59]. 2-BP is also capable of disturbing other PTMs such as myristoylation. Furthermore, high concentration and prolonged dealing time of 2-BP are often essential for S-palmitoylation inhibition, which causes intolerance in cell lines. Published studies have suggested that several cell lines, including, HeLa, HepG2, HEK293T and MDA-MB-231 exhibit cytotoxicity when dealt with 2-BP at concentrations of 20–80µM for 6–24 h [25, 187].

Cyanomethyl-myracrylamide (CMA)

In an effort to address the significant limitation associated with 2-BP, Saara-Anne Azizi et al. developed CMA, another pan-spectrum ZDHHCs inhibitor. Using an acrylamide warhead to replace α-halo FA of 2-BP could reduce toxicity and avoid inhibition of the S-palmitoylation eraser enzymes including APT1/2 [188]. Subsequent research reveals that CMA could block ZDHHC20 mediated S-palmitoylation of EGFR, consequently reduces the activation of PI3K and the abundance of transcription factor MYC [105]. CMA is also found to decrease CD36 S-palmitoylation, which contributes to reduce FA uptake and disturb lipid metabolism [189].

Disulfiram

Disulfiram, a FDA approved drug used for chronic alcoholism, has been recently found to inhibit NLRP3 inflammasome activation. Xu et al. reported that disulfiram inhibited S-palmitoylation of NLRP3 at Cys126, which is required for Golgi localization of NLRP3 inflammasome activation [190]. They also found that disulfiram could reduce NLRP3 S-palmitoylation when ZDHHC7 was knockdown, indicating that disulfiram also impacted other ZDHHC enzymes [190]. This finding suggests a mechanism that disulfiram targeted S-palmitoylation of NLRP3, providing a rationale of a safe FDA-approved drug for clinical application.

Therapeutic strategies targeting S-palmitoylation

Targeting protein palmitoylation holds significant therapeutic promise by precisely regulating protein localization, stability, and function. Drug resistance has become a huge challenge in tumor treatment. Using anti-CKAP4 antibody with third generation TKI shows stronger inhibition of tumor growth in lung cancer with EGFR mutations [191]. CKAP4 undergoes palmitoylation for its trafficking to exosomes, suggesting that inhibition of its palmitoylation may also reverse TKI resistance. Guo et al. revealed ZDHHC6-mediated MYD88 palmitoylation contributes to AML immunosuppression effect [127]. Blocking this modification may restores anti-tumor immunity. Besides, given the extensive involvement of palmitoylation in metabolic regulation, targeting this modification may represent a strategy to correct dysregulated metabolic states. Exploring these inhibitors on cellular functions and disease pathways will provide valuable insights into S-palmitoylation in health and disease. Further research into the development and optimization of S-palmitoylation inhibitors may lead to novel therapeutic strategies for treating conditions associated with aberrant S-palmitoylation protein.

Conclusions

S-palmitoylation, a key PTM involving palmitate attachment, plays a crucial role in diverse cellular processes. Recent methodological advances are enhancing our capacity to study S-palmitoylation and opening up new possibilities for its application. Notably, innovations in chemical biology have yielded tools for investigating S-palmitoylation, including clickable palmitate analogs, bioorthogonal chemistry, and proximity-based labeling strategies. These enable high-sensitivity visualization and quantification of S-palmitoylated proteins [192, 193]. Complementing these approaches, mass spectrometry-based proteomics is emerging as a powerful tool for global profiling of protein S-palmitoylation. Advances in mass spectrometry methods, such as acyl-biotin exchange and metabolic labeling, have facilitated the identification of S-palmitoylated proteins and the mapping of S-palmitoylation sites on a proteome-wide scale [194]. Moreover, real-time imaging techniques, such as fluorescence microscopy and bioluminescence resonance energy transfer (BRET), offer valuable insights into the spatiotemporal dynamics of S-palmitoylation in living cells [195, 196]. These novel approaches provide critical insights into the spatiotemporal dynamics of S-palmitoylation in living systems, revealing subcellular trafficking patterns.

Despite these advances, there are several research gaps targeting S-palmitoylation that need to be addressed. A primary challenge lies in determining the specificity and cellular dynamics of S-palmitoylation events. Identifying the exact residues of S-palmitoylation on specific proteins and elucidating the regulatory mechanisms involved remain areas of active investigation. While the importance of S-palmitoylation in protein localization and function is relatively well recognized, there is still much to learn about the specific functional consequences of S-palmitoylation for individual proteins. Understanding how it modulates protein stability, interactions, and signaling pathways is essential to decipher its biological significance. Despite evidence linking aberrant S-palmitoylation to diverse diseases, the precise mechanisms underlying these associations demand deeper exploration.

Further investigation to the role of S-palmitoylation in disease pathogenesis could uncover new diagnostic markers and therapeutic targets, expanding its clinical translation potential. Specifically, identifying disease-specific S-palmitoylation patterns can develop novel biomarker panels for early detection and monitoring of disease progression. Moreover, targeting S-palmitoylation pathways holds promise for the development of novel therapeutic interventions. Modulating protein S-palmitoylation levels with small molecule inhibitors could offer therapeutic strategies for diseases characterized by dysregulated S-palmitoylation. Critically, understanding the role of S-palmitoylation in disease pathogenesis may pave the way for personalized medicine approaches tailored to individual patients based on their S-palmitoylation profiles.

In conclusion, S-palmitoylation research is rapidly advancing, with ongoing efforts to address current research gaps, develop innovative detection methods, and explore potential clinical applications. Through overcoming current challenges and harnessing the potential of S-palmitoylation in disease diagnosis and treatment, we can pioneer new opportunities for advancing personalized medicine and improving patient care. This review has synthesized S-palmitoylation from basic mechanisms to disease implications, underscoring the imperative for continued research and highlighting transformative role of S-palmitoylation in biomedicine and clinical practice.

Abbreviations

ABHD

α/β hydrolase domain-containing protein

ACC

Acetyl-CoA carboxylase

acetyl-CoA

Acetyl-coenzyme A

ACOX1

Acyl-CoA oxidase-1

ACS

Acyl-CoA synthetase

AD

Alzheimer’s disease

AKAP150

A-kinase anchoring protein-150

AMI

Acute myocardial infarction

AML

Acute myeloid leukemia

AMPARs

α-amino-3-hydroxy-5-methyl-4-isox-azoleproprionic acid receptors

AMPKα1

Adenosine activated protein kinase alpha-1

APL

Acute promyelocytic leukemia

APTs

Acyl-protein thioesterases

AR

Androgen receptor

Atf3

Activating transcription factor-3

BACE1

β-site APP cleaving enzyme-1

BC

Breast cancer

BCa

Bladder cancer

BCL2

B-cell lymphoma-2

BCR

B cell receptor

BL

Burkitt lymphoma

BRET

Bioluminescence resonance energy transfer

CASP6

Caspase-6

CKAP4

Cytoskeleton-associated protein-4

CLDN3

Claudin-3

CLDN6

Claudin-6

CLL

Chronic lymphocytic leukemia

CMA

Cyanomethyl-myracrylamide

CPS1

Carbamoyl-phosphate synthetase-1

CRC

Colorectal cancer

CSCs

Cancer stem cells

DHHC

Asp-His-His-Cys

DPG

Aspartic acid-proline-glycine

eCB

Endocannabinoid

EGFR

Epidermal growth factor receptor

EP300

E1A binding protein P300

ER

Endoplasmic reticulum

ERα

Estrogen receptor-α

EZH2

Enhancer of zeste homolog

FA

Fatty acid

FASN

Fatty acid synthase

FGFR

Fibroblast growth factor receptor

GA

Golgi apparatus

GABAA

γ-aminobutyric acid sub-type A

GAD65

Glutamic acid decarboxylase-65

GBM

Glioblastoma

GC

Gastric carcinoma

GLUT

GCB-DLBCL, germinal center B-cell diffuse large B-cell lymphoma glucose transporter

GNA13

G protein subunit alpha-13

Golga7b

Golgin A7 family member B

GRIP1b

Glutamate receptor interacting protein-1

HA

Hyaluronic acid

HCC

Hepatocellular carcinoma

HD

Huntington disease

HIP14

Huntingtin interacting protein 14

HK1

Hexokinase-1

HSV

Herpes simplex virus

HTT

Huntingtin

IBD

Inflammatory bowel disease

IFNGR1

Interferon gamma receptor-1

IFITM3

Interferon-induced transmembrane protein-3

IGSF8

Immunoglobulin superfamily member-8

IRHOM2

Inactive rhomboid protein-2

ITGβ4

Integrin beta 4

LPS

Lipopolysaccharide

LRP6

LDL receptor-related protein-6

LYPLA

Lysophospholipase

MAP6

Microtubule-associated protein-6

MDH2

Malate dehydrogenase-2

MDSC

Myeloid-derived suppressor cells

NASH

Nonalcoholic steatohepatitis

NCX1

Na⁺/Ca²⁺ exchanger-1

NLRP3

Nucleotide-binding oligomerization domain leucine-rich repeat and pyrin-domain-containing-3

NMDARs

N-methyl-D-aspartate receptors

NOD2

Nucleotide oligomerization domain-like receptors 2

NSCLC

Non-small cell lung cancer

Nrf2

Nuclear factor erythroid 2-related factor-2

NTSR1

Neurotensin receptor-1

OC

Ovarian cancer

PA

Palmitic acid

PaCCT

Palmitoyl transferase conserved carboxy-terminal

PATs

Palmitoyl acyltransferases

PCa

Prostate cancer

PCSK9

Proprotein convertase subtilisin/kexin-9

PD

Parkinson’s disease

PDAC

Pancreatic ductal adenocarcinoma

PDE10A

Phosphodiesterase-10 A

PD-L1

Programmed cell death ligand-1

PI3K

Phosphatidylinositol 3

PLSCR1

Phospholipid scramblase-1

PML/RARα

Promyelocytic leukemia/retinoic acid receptor alpha

pm/mtEGFR

Plasm membrane and mitochondrion EGFR

PPARα

Peroxisome proliferator-activated receptor alpha

PPTs

Palmitoyl-protein thioesterases

PSD95

Postsynaptic density protein-95

PTM

Post-translational modification

PTP1B

Protein tyrosine phosphatase 1B

RCC

Renal cell carcinoma

ROS

Reactive oxygen species

SCM

Septic cardiomyopathy

SELENOK

Selenoprotein K

SLC1A3

Solute carrier family 1 member-3

SLE

Systemic lupus erythematosus

Smad3

SMAD family member-3

SNAP25

Synaptosome associated protein-25

SQSTM1

Sequestosome 1

SREBP1

Sterol regulatory element-binding protein-1

STAT3

Signal transducer and activator of transcription-3

STING

Stimulator of interferon genes

SYT1

Synaptotagmin-1

TBK1

TANK binding kinase-1

TCA cycle

Tricarboxylic acid cycle

TEAD

Transcriptional enhanced associate domain

TGF

Transforming growth factor

TGR5

Takeda G protein-coupled receptor-5

TKI

Tyrosine kinase inhibitor

TLR9

Toll-like receptor 9

TMD

Transmembrane domains

TTxE

Threonine-threonine-x-glutamic acid

UM

Uveal melanoma

VDAC2

Voltage dependent anion channel-2

YTHDF3

YTH domain-containing family protein-3

2-BP

2-bromopalmitate

Author contributions

Juanjuan Shang: wrote this manuscript, created figures and tables. Mei Ding and Xiangxiang Zhou: reviewed and revised the manuscript. Xiangxiang Zhou: provided direction and guidance throughout the preparation of the manuscript. All authors read and approved the final manuscript.

Funding

This work was supported by National Natural Science Foundation (No.82170189, No.81800194); Shandong Provincial Natural Science Foundation (ZR2021YQ51); Taishan Scholars Program of Shandong Province.

Data availability

No datasets were generated or analysed during the current study.

Declarations

Ethics approval and consent to participate

Not applicable.

Consent for publication

Not applicable.

Competing interests

The authors declare no competing interests.