Skip to main content
NCBI home page
Molecular Biology of the Cell logoLink to Molecular Biology of the Cell
. 2022 Apr 14;33(5):tp2. doi: 10.1091/mbc.E21-07-0363

An update on genetically encoded lipid biosensors

Gerald R V Hammond a,†,*, Morgan M C Ricci a,†, Claire C Weckerly a,†, Rachel C Wills a,†
Editor: William Bementb
PMCID: PMC9282013  PMID: 35420888

Abstract

Specific lipid species play central roles in cell biology. Their presence or enrichment in individual membranes can control properties or direct protein localization and/or activity. Therefore, probes to detect and observe these lipids in intact cells are essential tools in the cell biologist’s freezer box. Herein, we discuss genetically encoded lipid biosensors, which can be expressed as fluorescent protein fusions to track lipids in living cells. We provide a state-of-the-art list of the most widely available and reliable biosensors and highlight new probes (circa 2018–2021). Notably, we focus on advances in biosensors for phosphatidylinositol, phosphatidic acid, and PI 3-kinase lipid products.

INTRODUCTION

Lipids are fundamental building blocks of cellular life. Their amphiphilic nature makes them a keystone of bilayer membranes, as simply and elegantly illustrated by the double-tailed “tadpoles” of so many BioRender cartoons. Yet this deft simplicity belies the diversity of phospholipid, sphingolipid, and sterol species that make up biological membranes. The tightly crafted recipe of these lipids, with their unique shapes and charges, endows key functional properties on membranes: fluidity, curvature, and the capacity to selectively recruit or activate proteins are all regulated by lipids (Meer et al., 2008; Meer and Kroon, 2011; Balla, 2012). For this reason, cell biology demands approaches that can detect and enumerate membrane lipid compositions in their native cellular environment (Stahelin, 2009; Narwal et al., 2018; Dickson and Hille, 2019; Quinville et al., 2021).

This is where the genetically encoded lipid biosensors enter: these are typically lipid-binding domains from effector proteins or pathogen toxins, engineered to incorporate a tag for detection. Most conveniently, this involves fusion to a fluorescent protein for imaging in live cells. In this way, lipid biosensors can give information about the relative abundance, dynamics, and subcellular localization of lipids—in real time and in living cells. On the downside, the biosensors may be subject to biases in their localization, especially when not thoroughly characterized. We previously proposed two main criteria a biosensor should satisfy: 1) Is the biosensor selective for the lipid? This is typically determined in vitro. 2) Is the presence of the lipid both necessary and sufficient to localize the biosensor? This must be determined by modulation of lipids in the native cellular environment, and is often overlooked (Wills et al., 2018). Other caveats that must be considered include limitation to the detection of lipids in the outer plasma membrane or cytosolic membrane, because limits of fluorescence microscopy make interpreting localization in organelle lumens challenging. There are also extreme challenges to calibration, generally preventing quantification in terms of absolute lipid mass or mole fraction, though there have been technical tour-de-force studies that have done so (e.g., Liu et al., 2018). The strengths and weaknesses of lipid biosensors have already been explored in depth by ourselves and others (Balla et al., 2000; Lemmon, 2003; Maekawa and Fairn, 2014; Hammond and Balla, 2015; Wills et al., 2018). Suffice to say here, when it comes to genetically encoded lipid biosensors, a quote from Han Mi-nyeo, a character in the hit Netflix show Squid Game, sums it up: “I’m good at everything, except the things I can’t do.”

There have been many comprehensive reviews detailing currently available lipid biosensors (Stahelin, 2009; Kay et al., 2012; Maekawa and Fairn, 2014; Hammond and Balla, 2015; Narwal et al., 2018; Wills et al., 2018). We refer the reader to these resources for a comprehensive picture. Our goal here is to summarize a few notable recent advances and tools available for specific lipids. We also present an updated table (Table 1) showing some of the most widely used and (in our opinion) reliable genetically encoded lipid biosensors.

TABLE 1:

Current genetically encoded lipid biosensors for a variety of selective lipid species.

Lipid Biosensor Affinity Lipid specific? Cellular localization of lipid References
Lipid dependent? Lipid sufficient?
Chol D4-PFO + mutants 2–30 mol% ✔ ✔ ? Shimada et al., 2002; Maekawa and Fairn, 2015; Liu et al., 2016
SM Lysenin Kd ∼ 5 nM ✔ ✔ ? Yamaji et al., 1998; Kiyokawa et al., 2005; Abe et al., 2012
PA NES-PABD-spo20p51–91 (PASS) ? ✘ - binds PI(4,5)P2 and PIP3 weakly ✔ ? Zhang et al., 2014
NES-2xPABD-spo20p51–91 ? ✘ - suspected to bind PI(4,5)P2 and PIP3 weakly ✔ ? Bohdanowicz et al., 2013
α-Syn-N Kd ∼ 6.6 μM (18:1/18:1 PA) ✔ ✔ ? Yamada et al., 2020
PS C2-lactadherin Kd ∼ 0.5 µM ✔ ✔ ✔ Yeung et al., 2008; Maeda et al., 2013; Vecchio and Stahelin, 2018
DAG C1ab-PDK1 Ki (PDBu) ∼ 0.2 µM ✔ ✔ ✔ Chen et al., 2008; Kim et al., 2011
C1ab-PKCε Kd ∼ 10 nM ✔ ✔ ? Stahelin et al., 2005; Domart et al., 2012
PI BcPI-PLCH82A ? ✘ - binds to PC ✔ ✔ Pemberton et al., 2020
BcPI-PLCANH ? ✘ - binds DAG ✔ ✔ Pemberton et al., 2020
PI4P P4M-SidM Kd ∼ 1 µM or ∼18.2 nM FL ✔ ✔ ✔ Brombacher et al., 2009; Schoebel et al., 2010; Hammond et al., 2014
P4M-SidMx2 Kd < P4M-SidM ✔ ✔ ✔ Hammond et al., 2014; Levin et al., 2017
P4C-SidC Kd ∼ 250 nM ✔ ✔ ✔ Dolinsky et al., 2014; Weber et al., 2014; Zewe et al., 2018
N-PH-ORP5, N-PH-ORP8 Kd ∼ 5 µM for PI(4,5)P2 ✘ - binds PI4P and PI(4,5)P2 ✔ ✘ - requires PI(4,5)P2 Chung et al., 2015; Ghai et al., 2017; Sohn et al., 2018
PH-OSBP, PH-FAPP1 Kd ∼ 250 nM ✘ - binds PI(4,5)P2 ✔ ✘ - requires Arf1 Levine and Munro, 2002; Szentpetery et al., 2010; Lenoir et al., 2015
PI5P 3xPHD (ING2) ? ✘ - binds to PI3P ✔ ✔ Gozani et al., 2003; Pendaries et al., 2006
PI(4,5)P2 PH-PLCδ1 Kd ∼ 2 µM ✘ - binds to IP3∼20-fold more tightly than PI(4,5)P2 ✔ ✔ Garcia et al., 1995; Lemmon et al., 1995; Stauffer et al., 1998; Várnai and Balla, 1998; Hirose et al., 1999; Suh et al., 2006
PH-PLCδ4 Kd > PH-PLCδ1 ✘ - binds to IP3 ✔ ✔ Lee et al., 2004; Hammond and Balla, 2015
Tubbyc Kd > PH-PLCδ1 ✘ - binds PI(3,4)P2 and PI(3,4,5)P3 ✔ ✔ Quinn et al., 2008; Szentpetery et al., 2008; Halaszovich et al., 2009; Hammond and Balla, 2015
TubbycR332H Kd > Tubby ✘ - binds PI(3,4)P2 and PIP3 ✔ ? Quinn et al., 2008
ENTH/ANTH Kd ∼ 2 µM ✘ - binds to PIP3 ✔ ? Ford et al., 2001; Itoh et al., 2001; Yoon et al., 2011
PI3P FYVE-Hrsx2 Kd ∼ 2.5 µM ✔ ✔ ? Burd and Emr, 1998; Gaullier et al., 1998; Gillooly et al., 2000; Sankaran et al., 2001
FYVE-EEA1 Kd ∼ 45 nM ✔ ✔ ? Burd and Emr, 1998; Gaullier et al., 1998, 2000
PX-p40phox Kd ∼ 5 µM ✔ ✔ ? Bravo et al., 2001; Ellson et al., 2001; Kanai et al., 2001
PI(3,5)P2 ML1-Nx2 Kd ∼ 5.6 µM ✔ ✔/✘a ✔/✘a Li et al., 2013; Hammond et al., 2015
PI(3,4)P2 PH-TAPP1-CT Kd ∼ 80 nM ✔ ✔ ✔ Dowler et al., 2000; Thomas et al., 2001; Kimber et al., 2002; Marshall et al., 2002; Manna et al., 2007
eTapp1-PHc Kd ∼ 80 nM ✔ ✔ ✔ Liu et al., 2018
TAPP1-cPHx3 Kd > 80 nM ✔ ✔ ✔ Goulden et al., 2019
PIP3 PH-ARNO2G-I303Ex2 Kd ∼ 170 nM ✘ - binds IP4 ✔ ✔ Goulden et al., 2019
eMyoX-PHx2c Kd ∼ 33 nMb ✘ - binds IP4 ✔ ✔ Hokanson et al., 2006; Plantard et al., 2010; Lu et al., 2011; Liu et al., 2018
PH-Akt Kd ∼ 590 nM ✘ - binds PI(3,4)P2 and IP4 ✔ ? Frech et al., 1997; Watton and Downward, 1999; Manna et al., 2007
PH-Btk Kd ∼ 80 nM ✘ - binds IP4 ✔ ? Fukuda et al., 1996; Salim et al., 1996; Rameh et al., 1997; Kontos et al., 1998; Manna et al., 2007
PH-GRP1 (2G), PH-ARNO (2G) Kd ∼ 170 nM ✘ - binds IP4 ✔ ✘ - binds Arf/Arl Klarlund et al., 1997; Venkateswarlu et al., 1998; Gray et al., 1999; Cohen et al., 2007; Hofmann et al., 2007; Li et al., 2007; Manna et al., 2007
Open in a new tab

aThe accuracy of this probe is disputed.

bThe Kd value is derived from myosin-c tail and IP4 headgroup binding.

cRequires chemical ligation with a solvatochromic dye for optimal performance.

The phosphoinositide that we all forget about

There have been many iterations of biosensors for phosphoinositides, which are cardinal regulators of membrane function (Dickson and Hille, 2019). These lipids are all phosphorylated derivatives of a single parent lipid, phosphatidylinositol (PI). PI is a major lipid, approximately 10% of cellular phospholipids, with the derivatives being <1% (Vance, 2015), but a biosensor for PI itself had been lacking. As an abundant lipid, its distribution may be assumed to be ubiquitous. Yet, because PI is the key substrate for synthesis of the other phosphoinositides, its availability in given membranes is an important variable best not left to assumption.

Pemberton et al. recently created a PI biosensor utilizing the Bacillus cereus PI-specific phospholipase C (BcPI-PLC). BcPI-PLC was mutated in order to eliminate catalytic activity of the enzyme, yet retain the active site configuration that can accommodate the inositol headgroup (BcPI-PLCH82A). However, in vitro assays showed nonspecific BcPI-PLCH82A binding to PC-containing liposomes. To remove this nonspecific binding, two membrane-penetrating tyrosine residues were mutated to create the BcPI-PLCANH probe. It should be noted, though, that neither the BcPI-PLCH82A nor the BcPI-PLCANH probes are fully specific for PI in vitro, as the BcPI-PLCANH showed enhanced binding to liposomes containing DAG and PI (Pemberton et al., 2020). PI was shown to be necessary for membrane localization of both sensors, because depletion of PI by PI-PLC recruitment or AngII stimulation caused a decrease in membrane localization of the biosensors. The sufficiency of PI for biosensor recruitment was demonstrated when PI levels at the plasma membrane were increased with either pseudojanin-induced degradation of PI4P and PI(4,5)P2 to PI, or GSK-A1 inhibition of PI4KA-mediated conversion of PI to PI4P (Pemberton et al., 2020). Notably, the BcPI-PLCANH probe showed similar patterns of localization compared with BcPI-PLCH82A within cells. However, the BcPI-PLCANH showed more cytosolic localization than BcPI-PLCH82A, indicating that BcPI-PLCH82A may be a higher affinity probe for PI.

These probes revealed a surprising distribution of PI: an abundance at the endoplasmic reticulum (ER), peroxisomal, Golgi, and mitochondrial cytosolic leaflets, some on the endosomal network, but a notable absence at the plasma membrane at steady state (Pemberton et al., 2020). Satisfyingly, these findings were corroborated by additional approaches, including acute activation of PI-PLC or PI4Ks to generate diacylglycerol or PI4P from PI localized in specific membranes, which could be detected with other biosensors for these lipids (Pemberton et al., 2020; Zewe et al., 2020), and the trafficking of exogenously applied fluorescent PI (Zewe et al., 2020). Taken together, these results support a model where PI within the ER is transferred to the plasma membrane (PM) and then quickly converted into PI4P and PI(4,5)P2 to maintain homeostasis of these crucial PM phosphoinositide species.

The phospholipid backbone: phosphatidic acid

Phosphatidic acid (PA) is a crucial lipid, being both an intermediate in more complex phospholipid biosynthesis, and a second messenger molecule in diacylglycerol kinase (DGK) and phospholipase D (PLD) signaling pathways (Thakur et al., 2019). The most widely used biosensor is the phosphatidic acid biosensor with superior sensitivity (PASS) developed by Zhang et al. (2014). An added nuclear export sequence (NES) to the Spo20 phosphatidic acid–binding domain (PABD) prevented accumulation of PASS within the nucleus. This newly designed probe was able to show clear translocation to the PM after stimulation with phorbol-12-myristate-13-acetate, without having to overexpose images. However, the PASS did still retain some slight binding to PI(4,5)P2 and PIP3 within liposomes that the original Spo20 biosensor also showed (Zhang et al., 2014). A higher avidity, tandem dimer has also been developed (Bohdanowicz et al., 2013).

The usefulness of these PA biosensors has been recently corroborated by some new, ingenious tools, which have increased confidence in the accuracy of the Spo20-based PA lipid biosensors. An optogenetic bacterial PLD demonstrated that PA production in a variety of organelles is indeed sufficient to recruit PASS (Tei and Baskin, 2020). Additionally, click chemistry was used to label the products of PLD transphosphatidylation reactions as a proxy for PA, PLD’s endogenous product. This method showed in real time that active PLDs localize to the PM, ER, and Golgi, with slight localization on endosomes, lysosomes, and the mitochondria (Liang et al., 2019; Tei and Baskin, 2020).

Recent work has gone into characterizing the N-terminus of α-synuclein as a novel PA biosensor (Yamada et al., 2020). Using liposomes, this construct (α-Syn-N) was shown to be selective for PA as compared with other lipids. However, it also showed higher selectivity for 18:1/18:1 PA species, which could limit its use in endogenous systems where many different acyl chains are likely to occur, and the 18:1/18:1 species is rare (Lorent et al., 2020). Within Cos7 cells, the α-Syn-N biosensor was shown to be dependent on PA, as it colocalized with wild-type DGKs and PLD, but not when catalytically dead enzymes or inhibitors were used to prevent PA production. However, it is still not clear that this biosensor will be as sensitive as PASS when PA levels are modulated in a more physiological context (Yamada et al., 2020). Therefore, we still recommend the more robustly characterized Spo20-based PA biosensors.

Class I PI 3-kinase products: both of them

The class I PI 3-kinase pathway is a paramount regulator of growth in metazoa; it is often activated in cancer and other diseases (Fruman et al., 2017). Mechanistically, PI 3-kinase signaling operates through production of the lipid second messenger PIP3 by 3-OH phosphorylation of PI(4,5)P2. PIP3 can then be converted (to varying extents) into an additional signal, PI(3,4)P2, by 5-OH phosphatases (Malek et al., 2017). Both PIP3 and PI(3,4)P2 interact with effector proteins, which may be selective for one or both lipids (Hawkins and Stephens, 2016). Therefore, distinguishing these two lipids, and their sub­cellular localizations, is vital for delineating PI3K signaling at the cellular level.

The most popular biosensor for PI3K signaling is the lipid-binding pleckstrin homology (PH) domain from its most famous effector, Akt. Although often mistaken for a PIP3-biosensor, this domain actually binds to both PIP3 and PI(3,4)P2 (Manna et al., 2007; Ebner et al., 2017; Liu et al., 2018; Goulden et al., 2019). It is worth noting that the isolated PH domain, from all three isoforms of Akt(1–3), actually exhibits a preference for PI(3,4)P2, although this preference only holds true for Akt2 in the context of the full-length protein (Liu et al., 2018). Therefore, the Akt PH domain-based biosensors can be fine indicators of PI3K activity, but they report the convolution of PIP3 and PI(3,4)P2 signals.

Our lab has recently published a highly selective and sensitive PI(3,4)P2 biosensor, cPHx3, made of a tandem trimer of the C-terminal PH (cPH) domain from tandem PH-domain–containing protein 1 (TAPP1) fused to a NES and a fluorescent protein tag (Goulden et al., 2019). The improved sensitivity for PI(3,4)P2, derived from the high avidity of the tandem trimers, was evident when we detected the lipid’s synthesis after insulin stimulation, which had not previously been evident with lipid biosensors or many biochemical approaches (Goulden et al., 2019). Through an assortment of orthogonal manipulations in cells, we were also able to demonstrate that PI(3,4)P2 was both necessary and sufficient to drive cPHx3 localization in cells.

As an alternative to tandem arrays, Liu and colleagues improved the membrane binding of a single cPH domain by mutating a methionine to a membrane-penetrating tryptophan residue. This would undoubtedly improve the binding of a fluorescent protein conjugate. However, cysteine residues were also removed or inserted to produce a single site for chemical ligation of a solvatochromic dye, generating eTAPP1-cPH (Liu et al., 2018). This solvatochromic dye exhibits a spectral shift when inserted into the hydrophobic bilayer, permitting ratiometric imaging of the probe’s membrane association. When calibrated against known mole fractions of PI(3,4)P2 in liposomes, precise quantification of lipid concentration was realized (Liu et al., 2018). Therefore, precise spatiotemporal detection of PI(3,4)P2 is now possible, which when combined with recent advances in mass spectrometry detection of this lipid (Malek et al., 2017), will usher in a new era of understanding of this enigmatic lipid’s role in PI3K singling.

It is important to note that single PH domains from TAPP1 had previously been used as highly selective (but less sensitive) PI(3,4)P2 biosensors—but these came in two forms: one corresponding to the isolated cPH domain, and a second that includes the entire C-terminus of the protein. This C-terminal region contains a clathrin-binding domain, which biases the localization of the probe (Goulden et al., 2019). Therefore, it is critical to work with domains restricted to the isolated PH domains.

We also took advantage of the highly PIP3-selective 2G splice variant of the ARNO (also known as cytohesin-2) PH domain to make a high avidity, tandem dimer probe for this lipid (Cronin et al., 2004). We engineered an I303E mutation into each domain to disrupt a secondary binding site for Arl-family GTPases; this biosensor showed excellent selectivity for PIP3 in cells (Goulden et al., 2019). Liu et al. also engineered optimized membrane binding and solvatochromic dye–conjugated derivatives of the PIP3-selective MyosinX tandem PH domains, eMyoX-PH (Liu et al., 2018). Thus, there are now highly sensitive and selective PIP3 biosensors to accompany PI(3,4)P2 biosensors. These are included in Table 1. As noted in the table, a potential caveat to these sensors is their binding to soluble inositol tetrakisphosphate (IP4), the cognate headgroup of PIP3. This could potentially limit membrane translocation when PLC-mediated IP4 production is triggered in conjunction with PI3K. This is expected to be a more minor caveat for the dimeric probes, where local concentration of the lipid on the membrane will favor high avidity binding to the tandem PH domains.

CONCLUSION

Genetically encoded lipid biosensors continue to be a powerful and convenient tool to study lipid dynamics and function in cell biology. Here, we have focused on a brief refresher of the principles, and highlighted some of the newest biosensors that have appeared in the last 3 years. Given the recent trend from the last 3 years in biosensor development, it seems certain that new and improved probes are on the horizon, so we encourage the reader to continue keeping an eye open for the latest developments!

Acknowledgments

We apologize to colleagues whose work we have overlooked due to space considerations. This work was funded by National Institutes of Health Grant no. 1R35GM-119412-05.

Abbreviations used:

Chol

cholesterol

DAG

diacylglycerol

DGK

diacylglycerol kinase

ER

endoplasmic reticulum

IP3

inositol 1,4,5-trisphosphate

IP4

inositol 1,3,4,5-tetrakisphosphate

NES

nuclear export sequence

PA

phosphatidic acid

PABD

phosphatidic acid binding domain

PASS

phosphatidic acid biosensor with superior sensitivity

PC

phosphatidylcholine

PH

pleckstrin homology

PI

phosphatidylinositol

PI(3,4)P 2

phosphatidylinositol 3,4-bisphosphate

PI(3,5)P 2

phosphatidylinositol 3,5-bisphosphate

PI(4,5)P 2

phosphatidylinositol 4,5-bisphosphate

PI3K

phosphoinositide 3-kinase

PI3P

phosphatidylinositol 3-phosphate

PI4K

phosphatidylinositol 4-kinase

PI4P

phosphatidylinositol 4-phosphate

PI5P

phosphatidylinositol 5-phosphate

PIP 3

phosphatidylinositol 3,4,5-trisphosphate

PLC

phospholipase C

PLD

phospholipase D

PM

plasma membrane

PS

phosphatidylserine

SM

sphingomyelin

Footnotes

REFERENCES

  1. Abe M, Makino A, Hullin-Matsuda F, Kamijo K, Ohno-Iwashita Y, Hanada K, Mizuno H, Miyawaki A, Kobayashi T (2012). A role for sphingomyelin-rich lipid domains in the accumulation of phosphatidylinositol-4,5-bisphosphate to the cleavage furrow during cytokinesis. Mol Cell Biol 32, 1396–1407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Balla T (2012). Phosphoinositides: tiny lipids with giant impact on cell regulation. Physiol Rev 93, 1019–1137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Balla T, Bondeva T, Várnai P (2000). How accurately can we image inositol lipids in living cells? Trends Pharmacol Sci 21, 238–241. [DOI] [PubMed] [Google Scholar]
  4. Bohdanowicz M, Schlam D, Hermansson M, Rizzuti D, Fairn GD, Ueyama T, Somerharju P, Du G, Grinstein S (2013). Phosphatidic acid is required for the constitutive ruffling and macropinocytosis of phagocytes. Mol Biol Cell 24, 1700–1712. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bravo J, Karathanassis D, Pacold CM, Pacold ME, Ellson CD, Anderson KE, Butler PJ, Lavenir I, Perisic O, Hawkins PT, et al. (2001). The crystal structure of the PX domain from p40phox bound to phosphatidylinositol 3-phosphate. Mol Cell 8, 829–839. [DOI] [PubMed] [Google Scholar]
  6. Brombacher E, Urwyler S, Ragaz C, Weber SS, Kami K, Overduin M, Hilbi H (2009). Rab1 guanine nucleotide exchange factor SidM is a major phosphatidylinositol 4-phosphate-binding effector protein of Legionella pneumophila. J Biol Chem 284, 4846–4856. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Burd CG, Emr SD (1998). Phosphatidylinositol(3)-phosphate signaling mediated by specific binding to RING FYVE domains. Mol Cell 2, 157–162. [DOI] [PubMed] [Google Scholar]
  8. Chen J, Deng F, Li J, Wang QJ (2008). Selective binding of phorbol esters and diacylglycerol by individual C1 domains of the PKD family. Biochem J 411, 333–342. [DOI] [PubMed] [Google Scholar]
  9. Chung J, Torta F, Masai K, Lucast L, Czapla H, Tanner LB, Narayanaswamy P, Wenk MR, Nakatsu F, Camilli PD (2015). PI4P/phosphatidylserine countertransport at ORP5- and ORP8-mediated ER–plasma membrane contacts. Science 349, 428–432. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cohen LA, Honda A, Varnai P, Brown FD, Balla T, Donaldson JG (2007). Active Arf6 recruits ARNO/cytohesin GEFs to the PM by binding their PH domains. Mol Biol Cell 18, 2244–2253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cronin TC, DiNitto JP, Czech MP, Lambright DG (2004). Structural determinants of phosphoinositide selectivity in splice variants of Grp1 family PH domains. EMBO J 23, 3711–3720. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dickson EJ, Hille B (2019). Understanding phosphoinositides: rare, dynamic, and essential membrane phospholipids. Biochem J 476, 1–23. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dolinsky S, Haneburger I, Cichy A, Hannemann M, Itzen A, Hilbi H (2014). The Legionella longbeachae Icm/dot substrate SidC selectively binds phosphatidylinositol 4-phosphate with nanomolar affinity and promotes pathogen vacuole-endoplasmic reticulum interactions. Infect Immun 82, 4021–4033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Domart MC, Hobday TM, Peddie CJ, Chung GH, Wang A, Yeh K, Jethwa N, Zhang Q, Wakelam MJ, Woscholski R, et al. (2012). Acute manipulation of diacylglycerol reveals roles in nuclear envelope assembly and endoplasmic reticulum morphology. PLoS One 7, e51150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Dowler S, Currie RA, Campbell DG, Deak M, Kular G, Downes CP, Alessi DR (2000). Identification of pleckstrin-homology-domain-containing proteins with novel phosphoinositide-binding specificities. Biochem J 351, 19–31. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ebner M, Lucˇic´ I, Leonard TA, Yudushkin I (2017). PI(3,4,5)P3 engagement restricts Akt activity to cellular membranes. Mol Cell 65, 416–431.e6. [DOI] [PubMed] [Google Scholar]
  17. Ellson CD, Gobert-Gosse S, Anderson KE, Davidson K, Erdjument-Bromage H, Tempst P, Thuring JW, Cooper MA, Lim ZY, Holmes AB, et al. (2001). PtdIns(3)P regulates the neutrophil oxidase complex by binding to the PX domain of p40phox. Nat Cell Biol 3, 679–682. [DOI] [PubMed] [Google Scholar]
  18. Ford MGJ, Pearse BMF, Higgins MK, Vallis Y, Owen DJ, Gibson A, Hopkins CR, Evans PR, McMahon HT (2001). Simultaneous binding of PtdIns(4,5)P2 and clathrin by AP180 in the nucleation of clathrin lattices on membranes. Science 291, 1051–1055. [DOI] [PubMed] [Google Scholar]
  19. Frech M, Andjelkovic M, Ingley E, Reddy KK, Falck JR, Hemmings BA (1997). High affinity binding of inositol phosphates and phosphoinositides to the pleckstrin homology domain of RAC/protein kinase B and their influence on kinase activity. J Biol Chem 272, 8474–8481. [DOI] [PubMed] [Google Scholar]
  20. Fruman DA, Chiu H, Hopkins BD, Bagrodia S, Cantley LC, Abraham RT (2017). The PI3K pathway in human disease. Cell 170, 605–635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Fukuda M, Kojima T, Kabayama H, Mikoshiba K (1996). Mutation of the pleckstrin homology domain of Bruton’s tyrosine kinase in immunodeficiency impaired inositol 1,3,4,5-tetrakisphosphate binding capacity. J Biol Chem 271, 30303–30306. [DOI] [PubMed] [Google Scholar]
  22. Garcia P, Gupta R, Shah S, Morris AJ, Rudge SA, Scarlata S, Petrova V, McLaughlin S, Rebecchi MJ (1995). The pleckstrin homology domain of phospholipase C-delta 1 binds with high affinity to phosphatidylinositol 4,5-bisphosphate in bilayer membranes. Biochem-US 34, 16228–16234. [DOI] [PubMed] [Google Scholar]
  23. Gaullier J-M, Rønning E, Gillooly DJ, Stenmark H (2000). Interaction of the EEA1 FYVE finger with phosphatidylinositol 3-phosphate and early endosomes. role of conserved residues. J Biol Chem 275, 24595–24600. [DOI] [PubMed] [Google Scholar]
  24. Gaullier J-M, Simonsen A, D’Arrigo A, Bremnes B, Stenmark H, Aasland R (1998). FYVE fingers bind PtdIns(3)P. Nature 394, 432–433. [DOI] [PubMed] [Google Scholar]
  25. Ghai R, Du X, Wang H, Dong J, Ferguson C, Brown AJ, Parton RG, Wu J-W, Yang H (2017). ORP5 and ORP8 bind phosphatidylinositol-4, 5-biphosphate (PtdIns(4,5)P2) and regulate its level at the plasma membrane. Nat Commun 8, 757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Gillooly DJ, Morrow IC, Lindsay M, Gould R, Bryant NJ, Gaullier J, Parton RG, Stenmark H (2000). Localization of phosphatidylinositol 3-phosphate in yeast and mammalian cells. EMBO J 19, 4577–4588. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Goulden BD, Pacheco J, Dull A, Zewe JP, Deiters A, Hammond GRV (2019). A high-avidity biosensor reveals plasma membrane PI(3,4)P2 is predominantly a class I PI3K signaling product. J Cell Biol 218, 1066–1079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Gozani O, Karuman P, Jones DR, Ivanov D, Cha J, Lugovskoy AA, Baird CL, Zhu H, Field SJ, Lessnick SL, et al. (2003). The PHD finger of the chromatin-associated protein ING2 functions as a nuclear phosphoinositide receptor. Cell 114, 99–111. [DOI] [PubMed] [Google Scholar]
  29. Gray A, Vanderkaay J, Downes CP (1999). The pleckstrin homology domains of protein kinase B and GRP1 (general receptor for phosphoinositides-1) are sensitive and selective probes for the cellular detection of phosphatidylinositol 3,4-bisphosphate and/or phosphatidylinositol 3,4,5-trisphosphate in vivo. Biochem J 344, 929–936. [PMC free article] [PubMed] [Google Scholar]
  30. Halaszovich CR, Schreiber DN, Oliver D (2009). Ci-VSP is a depolarization-activated phosphatidylinositol-4,5-bisphosphate and phosphatidylinositol-3,4,5-trisphosphate 5′-phosphatase. J Biol Chem 284, 2106–2113. [DOI] [PubMed] [Google Scholar]
  31. Hammond GRV, Balla T (2015). Polyphosphoinositide binding domains: key to inositol lipid biology. Biochim Biophys Acta 1851, 746–758. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Hammond GRV, Machner MP, Balla T (2014). A novel probe for phosphatidylinositol 4-phosphate reveals multiple pools beyond the Golgi. J Cell Biol 205, 113–126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Hammond GRV, Takasuga S, Sasaki T, Balla T (2015). The ML1Nx2 phosphatidylinositol 3,5-bisphosphate probe shows poor selectivity in cells. PLoS One 10, e0139957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Hawkins PT, Stephens LR (2016). Emerging evidence of signalling roles for PI(3,4)P2 in class I and II PI3K-regulated pathways. Biochem Soc T 44, 307–314. [DOI] [PubMed] [Google Scholar]
  35. Hirose K, Kadowaki S, Tanabe M, Takeshima H, Iino M (1999). Spatiotemporal dynamics of inositol 1,4,5-trisphosphate that underlies complex Ca2+ mobilization patterns. Science 284, 1527–1530. [DOI] [PubMed] [Google Scholar]
  36. Hofmann I, Thompson A, Sanderson CM, Munro S (2007). The Arl4 family of small G proteins can recruit the cytohesin Arf6 exchange factors to the plasma membrane. Curr Biol 17, 711–716. [DOI] [PubMed] [Google Scholar]
  37. Hokanson ED, Laakso MJ, Lin T, Sept D, Ostap ME (2006). Myo1c binds phosphoinositides through a putative pleckstrin homology domain. MBoC 17, 4856–4865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Itoh T, Koshiba S, Kigawa T, Kikuchi A, Yokoyama S, Takenawa T (2001). Role of the ENTH domain in phosphatidylinositol-4,5-bisphosphate binding and endocytosis. Science 291, 1047–1051. [DOI] [PubMed] [Google Scholar]
  39. Kanai F, Liu H, Field SJ, Akbary H, Matsuo T, Brown GE, Cantley LC, Yaffe MB (2001). The PX domains of p47phox and p40phox bind to lipid products of PI(3)K. Nat Cell Biol 3, 675–678. [DOI] [PubMed] [Google Scholar]
  40. Kay JG, Koivusalo M, Ma X, Wohland T, Grinstein S (2012). Phosphatidylserine dynamics in cellular membranes. Mol Biol Cell 23, 2198–2212. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Kim YJ, Guzman-Hernandez ML, Balla T (2011). A highly dynamic ER-derived phosphatidylinositol-synthesizing organelle supplies phosphoinositides to cellular membranes. Dev Cell 21, 813–824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Kimber WA, Watt S, Javier RT, Gray A, Downes CP, Lucocq JM, Alessi DR (2002). Evidence that the tandem-pleckstrin-homology-domain-containing protein TAPP1 interacts with Ptd(3,4)P2 and the multi-PDZ-domain-containing protein MUPP1 in vivo. Biochem J 361, 525–536. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Kiyokawa E, Baba T, Otsuka N, Makino A, Ohno S, Kobayashi T (2005). Spatial and functional heterogeneity of sphingolipid-rich membrane domains. J Biol Chem 280, 24072–24084. [DOI] [PubMed] [Google Scholar]
  44. Klarlund JK, Guilherme A, Holik JJ, Virbasius JV, Chawla A, Czech MP (1997). Signaling by phosphoinositide-3,4,5-trisphosphate through proteins containing pleckstrin and Sec7 homology domains. Science 275, 1927–1930. [DOI] [PubMed] [Google Scholar]
  45. Kontos CD, Stauffer TP, Yang W-P, York JD, Huang L, Blanar MA, Meyer T, Peters KG (1998). Tyrosine 1101 of Tie2 is the major site of association of p85 and is required for activation of phosphatidylinositol 3-kinase and Akt. Mol Cell Biol 18, 4131–4140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Lee SB, Várnai P, Balla A, Jalink K, Rhee S-G, Balla T (2004). The pleckstrin homology domain of phosphoinositide-specific phospholipase Cδ4 is not a critical determinant of the membrane localization of the enzyme. J Biol Chem 279, 24362–24371. [DOI] [PubMed] [Google Scholar]
  47. Lemmon MA (2003). Phosphoinositide recognition domains. Traffic 4, 201–213. [DOI] [PubMed] [Google Scholar]
  48. Lemmon MA, Ferguson KM, O’Brien R, Sigler PB, Schlessinger J (1995). Specific and high-affinity binding of inositol phosphates to an isolated pleckstrin homology domain. Proc Natl Acad Sci USA 92, 10472–10476. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Lenoir M, Grzybek M, Majkowski M, Rajesh S, Kaur J, Whittaker SB-M, Coskun Ü, Overduin M (2015). Structural basis of dynamic membrane recognition by trans-Golgi network specific FAPP proteins. J Mol Biol 427, 966–981. [DOI] [PubMed] [Google Scholar]
  50. Levin R, Hammond GRV, Balla T, Camilli PD, Fairn GD, Grinstein S (2017). Multiphasic dynamics of phosphatidylinositol 4-phosphate during phagocytosis. Mol Biol Cell 28, 128–140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Levine TP, Munro S (2002). Targeting of Golgi-specific pleckstrin homology domains involves both PtdIns 4-kinase-dependent and -independent components. Curr Biol 12, 695–704. [DOI] [PubMed] [Google Scholar]
  52. Li C-C, Chiang T-C, Wu T-S, Pacheco-Rodriguez G, Moss J, Lee F-Js (2007). ARL4D recruits cytohesin-2/ARNO to modulate actin remodeling. MBoC 18, 4420–4437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Li X, Wang X, Zhang X, Zhao M, Tsang WL, Zhang Y, Yau RGW, Weisman LS, Xu H (2013). Genetically encoded fluorescent probe to visualize intracellular phosphatidylinositol 3,5-bisphosphate localization and dynamics. Proc Natl Acad Sci USA 110, 21165–21170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Liang D, Wu K, Tei R, Bumpus TW, Ye J, Baskin JM (2019). A real-time, click chemistry imaging approach reveals stimulus-specific subcellular locations of phospholipase D activity. Proc Natl Acad Sci USA 116, 15453–15462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Liu SL, Sheng R, Jung JH, Wang L, Stec E, O’Connor MJ, Song S, Bikkavilli RK, Winn RA, Lee D, et al. (2016). Orthogonal lipid sensors identify transbilayer asymmetry of plasma membrane cholesterol. Nat Chem Biol 13, 268–274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Liu SL, Wang ZG, Hu Y, Xin Y, Singaram I, Gorai S, Zhou X, Shim Y, Min JH, Gong LW, et al. (2018). Quantitative lipid imaging reveals a new signaling function of phosphatidylinositol-3,4-bisphophate: isoform- and site-specific activation of Akt. Mol Cell 71, 1092–1104.e5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Lorent J, Levental K, Ganesan L, Rivera-Longsworth G, Sezgin E, Doktorova M, Lyman E, Levental I (2020). Plasma membranes are asymmetric in lipid unsaturation, packing, and protein shape. Nat Chem Biol 16, 644–652. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Lu Q, Yu J, Yan J, Wei Z, Zhang M (2011). Structural basis of the myosin X PH1N-PH2-PH1C tandem as a specific and acute cellular PI(3,4,5)P3 sensor. Mol Biol Cell 22, 4268–4278. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Maeda K, Anand K, Chiapparino A, Kumar A, Poletto M, Kaksonen M, Gavin A-C (2013). Interactome map uncovers phosphatidylserine transport by oxysterol-binding proteins. Nature 501, 257–261. [DOI] [PubMed] [Google Scholar]
  60. Maekawa M, Fairn GD (2014). Molecular probes to visualize the location, organization and dynamics of lipids. J Cell Sci 127, 4801–4812. [DOI] [PubMed] [Google Scholar]
  61. Maekawa M, Fairn GD (2015). Complementary probes reveal that phosphatidylserine is required for the proper transbilayer distribution of cholesterol. J Cell Sci 128, 1422–1433. [DOI] [PubMed] [Google Scholar]
  62. Malek M, Kielkowska A, Chessa T, Anderson KE, Barneda D, Pir P, Nakanishi H, Eguchi S, Koizumi A, Sasaki J, et al. (2017). PTEN regulates PI(3,4)P2 signaling downstream of class I PI3K. Mol Cell 68, 566–580.e10. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Manna D, Albanese A, Park WS, Cho W (2007). Mechanistic basis of differential cellular responses of phosphatidylinositol 3,4-bisphosphate- and phosphatidylinositol 3,4,5-trisphosphate-binding pleckstrin homology domains. J Biol Chem 282, 32093–32105. [DOI] [PubMed] [Google Scholar]
  64. Marshall AJ, Krahn AK, Ma K, Duronio V, Hou S (2002). TAPP1 and TAPP2 are targets of phosphatidylinositol 3-kinase signaling in B cells: sustained plasma membrane recruitment triggered by the B-cell antigen receptor. Mol Cell Biol 22, 5479–5491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  65. Meer G, de Kroon A (2011). Lipid map of the mammalian cell. J Cell Sci 124, 5–8. [DOI] [PubMed] [Google Scholar]
  66. Meer G, Voelker DR, Feigenson GW (2008). Membrane lipids: where they are and how they behave. Nat Rev Mol Cell Biol 9, 112–124. [DOI] [PMC free article] [PubMed] [Google Scholar]
  67. Narwal V, Deswal R, Batra B, Kalra V, Hooda R, Sharma M, Rana JS (2018). Cholesterol biosensors: a review. Steroids 143, 6–17. [DOI] [PubMed] [Google Scholar]
  68. Pemberton JG, Kim YJ, Humpolickova J, Eisenreichova A, Sengupta N, Toth DJ, Boura E, Balla T (2020). Defining the subcellular distribution and metabolic channeling of phosphatidylinositol. J Cell Biol 219, e201906130. [DOI] [PMC free article] [PubMed] [Google Scholar]
  69. Pendaries C, Tronchère H, Arbibe L, Mounier J, Gozani O, Cantley L, Fry MJ, Gaits-Iacovoni F, Sansonetti PJ, Payrastre B (2006). PtdIns(5)P activates the host cell PI3-kinase/Akt pathway during Shigella flexneri infection. EMBO J 25, 1024–1034. [DOI] [PMC free article] [PubMed] [Google Scholar]
  70. Plantard L, Arjonen A, Lock JG, Nurani G, Ivaska J, Strömblad S (2010). PtdIns(3,4,5)P3 is a regulator of myosin-X localization and filopodia formation. J Cell Sci 123, 3525–3534. [DOI] [PubMed] [Google Scholar]
  71. Quinn KV, Behe P, Tinker A (2008). Monitoring changes in membrane phosphatidylinositol 4,5-bisphosphate in living cells using a domain from the transcription factor tubby. J Physiol 586, 2855–2871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  72. Quinville BM, Deschenes NM, Ryckman AE, Walia JS (2021). A comprehensive review: sphingolipid metabolism and implications of disruption in sphingolipid homeostasis. Int J Mol Sci 22, 5793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  73. Rameh LE, Arvidsson AK, Carraway KL 3rd, Couvillon AD, Rathbun G, Crompton A, VanRenterghem B, Czech MP, Ravichandran KS, Burakoff SJ, et al. (1997). A comparative analysis of the phosphoinositide binding specificity of pleckstrin homology domains. J Biol Chem 272, 22059–22066. [DOI] [PubMed] [Google Scholar]
  74. Salim K, Bottomley MJ, Querfurth E, Zvelebil MJ, Gout I, Scaife R, Margolis RL, Gigg R, Smith CI, Driscoll PC, et al. (1996). Distinct specificity in the recognition of phosphoinositides by the pleckstrin homology domains of dynamin and Bruton’s tyrosine kinase. EMBO J 15, 6241–6250. [PMC free article] [PubMed] [Google Scholar]
  75. Sankaran VG, Klein DE, Sachdeva MM, Lemmon MA (2001). High-affinity binding of a FYVE domain to phosphatidylinositol 3-phosphate requires intact phospholipid but not FYVE domain oligomerization. Biochem-US 40, 8581–8587. [DOI] [PubMed] [Google Scholar]
  76. Schoebel S, Blankenfeldt W, Goody RS, Itzen A (2010). High-affinity binding of phosphatidylinositol 4-phosphate by Legionella pneumophila DrrA. EMBO Rep 11, 598–604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  77. Shimada Y, Maruya M, Iwashita S, Ohno-Iwashita Y (2002). The C-terminal domain of perfringolysin O is an essential cholesterol-binding unit targeting to cholesterol-rich microdomains. Eur J Biochem 269, 6195–6203. [DOI] [PubMed] [Google Scholar]
  78. Sohn M, Korzeniowski M, Zewe JP, Wills RC, Hammond G, Humpolickova J, Vrzal L, Chalupska D, Veverka V, Fairn GD, et al. (2018). PI(4,5)P2 controls plasma membrane PI4P and PS levels via ORP5/8 recruitment to ER–PM contact sites. J Cell Biol 217, 1797–1813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  79. Stahelin RV (2009). Lipid binding domains: more than simple lipid effectors. J Lipid Res 50, S299–S304. [DOI] [PMC free article] [PubMed] [Google Scholar]
  80. Stahelin RV, Digman MA, Medkova M, Ananthanarayanan B, Melowic HR, Rafter JD, Cho W (2005). Diacylglycerol-induced membrane targeting and activation of protein kinase Cε: mechanistic differences between protein kinases Cδ and Cε. J Biol Chem 280, 19784–19793. [DOI] [PubMed] [Google Scholar]
  81. Stauffer TP, Ahn S, Meyer T (1998). Receptor-induced transient reduction in plasma membrane PtdIns(4,5)P2 concentration monitored in living cells. Curr Biol 8, 343–346. [DOI] [PubMed] [Google Scholar]
  82. Suh B-C, Inoue T, Meyer T, Hille B (2006). Rapid chemically induced changes of PtdIns(4,5)P2 gate KCNQ ion channels. Science 314, 1454–1457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  83. Szentpetery Z, Balla A, Kim YJ, Lemmon MA, Balla T (2008). Live cell imaging with protein domains capable of recognizing phosphatidylinositol 4,5-bisphosphate; a comparative study. BMC Cell Biol 10, 67. [DOI] [PMC free article] [PubMed] [Google Scholar]
  84. Szentpetery Z, Várnai P, Balla T (2010). Acute manipulation of Golgi phosphoinositides to assess their importance in cellular trafficking and signaling. Proc Natl Acad Sci USA 107, 8225–8230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  85. Tei R, Baskin JM (2020). Spatiotemporal control of phosphatidic acid signaling with optogenetic, engineered phospholipase Ds. J Cell Biol 219, e201907013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  86. Thakur R, Naik A, Panda A, Raghu P (2019). Regulation of membrane turnover by phosphatidic acid: cellular functions and disease implications. Front Cell Dev Biol 7, 83. [DOI] [PMC free article] [PubMed] [Google Scholar]
  87. Thomas CC, Dowler S, Deak M, Alessi DR, Aalten DM (2001). Crystal structure of the phosphatidylinositol 3,4-bisphosphate-binding pleckstrin homology (PH) domain of tandem PH-domain-containing protein 1 (TAPP1): molecular basis of lipid specificity. Biochem J 358, 287–294. [DOI] [PMC free article] [PubMed] [Google Scholar]
  88. Vance JE (2015). Phospholipid synthesis and transport in mammalian cells. Traffic 16, 1–18. [DOI] [PubMed] [Google Scholar]
  89. Várnai P, Balla T (1998). Visualization of phosphoinositides that bind pleckstrin homology domains: calcium- and agonist-induced dynamic changes and relationship to Myo-[3H]inositol-labeled phosphoinositide pools. J Cell Biol 143, 501–510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  90. Vecchio KD, Stahelin RV (2018). Investigation of the phosphatidylserine binding properties of the lipid biosensor, Lactadherin C2 (LactC2), in different membrane environments. J Bioenerg Biomembr 50, 1–10. [DOI] [PMC free article] [PubMed] [Google Scholar]
  91. Venkateswarlu K, Oatey PB, Tavaré JM, Cullen PJ (1998). Insulin-dependent translocation of ARNO to the plasma membrane of adipocytes requires phosphatidylinositol 3-kinase. Curr Biol 8, 463–466. [DOI] [PubMed] [Google Scholar]
  92. Watton SJ, Downward J (1999). Akt/PKB localisation and 3′ phosphoinositide generation at sites of epithelial cell–matrix and cell–cell interaction. Curr Biol 9, 433–436. [DOI] [PubMed] [Google Scholar]
  93. Weber S, Wagner M, Hilbi H (2014). Live-cell imaging of phosphoinositide dynamics and membrane architecture during Legionella infection. mBio 5, e00839-13. [DOI] [PMC free article] [PubMed] [Google Scholar]
  94. Wills RC, Goulden BD, Hammond GRV (2018). Genetically encoded lipid biosensors. Mol Biol Cell 29, 1526–1532. [DOI] [PMC free article] [PubMed] [Google Scholar]
  95. Yamada H, Mizuno S, Honda S, Takahashi D, Sakane F (2020). Characterization of α-synuclein N-terminal domain as a novel cellular phosphatidic acid sensor. Febs J 287, 2212–2234. [DOI] [PubMed] [Google Scholar]
  96. Yamaji A, Sekizawa Y, Emoto K, Sakuraba H, Inoue K, Kobayashi H, Umeda M (1998). Lysenin, a novel sphingomyelin-specific binding protein. J Biol Chem 273, P5300–P5306. [DOI] [PubMed] [Google Scholar]
  97. Yeung T, Gilbert GE, Shi J, Silvius J, Kapus A, Grinstein S (2008). Membrane phosphatidylserine regulates surface charge and protein localization. Science 319, 210–213. [DOI] [PubMed] [Google Scholar]
  98. Yoon Y, Lee PJ, Kurilova S, Cho W (2011). In situ quantitative imaging of cellular lipids using molecular sensors. Nat Chem 3, 868–874. [DOI] [PMC free article] [PubMed] [Google Scholar]
  99. Zewe JP, Miller AM, Sangappa S, Wills RC, Goulden BD, Hammond GRV (2020). Probing the subcellular distribution of phosphatidylinositol reveals a surprising lack at the plasma membrane. J Cell Biol 219, e201906127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  100. Zewe JP, Wills RC, Sangappa S, Goulden BD, Hammond GR (2018). SAC1 degrades its lipid substrate PtdIns4P in the endoplasmic reticulum to maintain a steep chemical gradient with donor membranes. Elife 7, e35588. [DOI] [PMC free article] [PubMed] [Google Scholar]
  101. Zhang F, Wang Z, Lu M, Yonekubo Y, Liang X, Zhang Y, Wu P, Zhou Y, Grinstein S, Hancock JF, et al. (2014). Temporal production of the signaling lipid phosphatidic acid by phospholipase D2 determines the output of extracellular signal-regulated kinase signaling in cancer cells. Mol Cell Biol 34, 84–95. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular Biology of the Cell are provided here courtesy of American Society for Cell Biology

RESOURCES