Appendix 1—key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background RH (Toxoplasma gondii) | TIR1 | PMID:28465425 | RH/TIR1/∆KU80/∆HXGPRT. Mycoplasma negative. | |
| Strain, strain background RH (T. gondii) | CDPK1-AID | doi:https://doi.org/10.1101/2022.07.19.500742 | RH/TIR1/∆KU80/∆HXGPRT/CDPK1-V5-mNeonGreen-mAID-Ty. Mycoplasma negative. | |
| Strain, strain background RH (T. gondii) | iKD CDPK1 | This paper | TGGT1_301440 | RH/TIR1/∆KU80/∆HXGPRT/CDPK1-mAID-Myc/HXGPRT. Mycoplasma negative. |
| Strain, strain background RH (T. gondii) | cWT CDPK1 | This paper | TGGT1_301440 | RH/TIR1/∆KU80/∆HXGPRT/CDPK1-mAID-Myc/HXGPRT/CDPK1-HA-T2A-GFP. Mycoplasma negative. |
| Strain, strain background RH (T. gondii) | cMut CDPK1 | This paper | TGGT1_301440 | RH/TIR1/∆KU80/∆HXGPRT/CDPK1-mAID-Myc/HXGPRT/CDPK1(G2A)-HA-T2A-mCherry. Mycoplasma negative. |
| Strain, strain background RH (T. gondii) | ∆ku80∆hxgprt YFP | PMID:32618271 | RH/∆KU80/∆HXGPRT/YFP. Mycoplasma negative. | |
| Strain, strain background RH (T. gondii) | CDPK1G | PMID:19218426 | RH/∆KU80/∆HXGPRT. Mycoplasma negative. | |
| Strain, strain background RH (T. gondii) | CDPK1M | PMID:23149386 | TGGT1_301440 | RH/∆KU80/∆HXGPRT/CDPK1(G128M)-Myc. Mycoplasma negative. |
| Strain, strain background RH (T. gondii) | AID-HOOK | This paper | TGGT1_289100 | RH/TIR1/∆KU80/∆HXGPRT/HA-mAID-HOOK. Mycoplasma negative. |
| Strain, strain background RH (T. gondii) | CDPK1-AID/HOOK-3xHA | This paper | TGGT1_289100 | RH/TIR1/∆KU80/∆HXGPRT/CDPK1-V5-mNeonGreen-mAID-Ty/HOOK-V5-3HA/HXGPRT. Mycoplasma negative. |
| Strain, strain background RH (T. gondii) | FTS-AID | This paper | TGGT1_264050 | RH/TIR1/∆KU80/∆HXGPRT/FTS-V5-mAID-HA/HXGPRT. Mycoplasma negative. |
| Strain, strain background RH (T. gondii) | 306920-3xHA | This paper | TGGT1_306920 | RH/TIR1/∆KU80/∆HXGPRT/306920-V5-3HA/HXGPRT. Mycoplasma negative. |
| Strain, strain background RH (T. gondii) | FTS-3xHA | This paper | TGGT1_264050 | RH/TIR1/∆KU80/∆HXGPRT/FTS-V5-3HA/HXGPRT. Mycoplasma negative. |
| Strain, strain background RH (T. gondii) | FTS-TurboID | This paper | TGGT1_264050 | RH/TIR1/∆KU80/∆HXGPRT/FTS-TurboID-Ty/DHFR. Mycoplasma negative. |
| Strain, strain background RH (T. gondii) | Cytosolic mNG-TurboID | This paper | RH/TIR1/∆KU80/∆HXGPRT/pMIC2-mNeonGreen-TurboID-Ty. Mycoplasma negative. | |
| Strain, strain background RH (T. gondii) | CLAMP-mNG | PMID:27594426 | TGGT1_265790 | RH/∆KU80/∆HXGPRT/CLAMP-mNeonGreen. Mycoplasma negative. |
| Strain, strain background RH (T. gondii) | TIR1/CLAMP-mNG | This paper | TGGT1_265790 | RH/TIR1/∆KU80/∆HXGPRT/CLAMP-mNeonGreen. Mycoplasma negative. |
| Strain, strain background RH (T. gondii) | TIR1/CLAMP-mNG/AID-HOOK | This paper | TGGT1_265790 | RH/TIR1/∆KU80/∆HXGPRT/HA-mAID-HOOK/CLAMP-mNeonGreen. Mycoplasma negative. |
| Cell line (Homo sapiens) | Human Foreskin Fibroblasts (HFFs) | ATCC | SCRC-1041 | Mycoplasma negative. |
| Antibody | Mouse polyclonal anti-CDPK1 | PMID:28246362 | Provided by Matthew Child and Matt Bogyo. WB (1:3000). Only used in Figure 2. | |
| Antibody | Mouse monoclonal clone 4A6 anti-Myc | Millipore | Cat# 05-724, RRID:AB_11211891 | WB (1:1000). |
| Antibody | Rat monoclonal clone 3F10 anti-HA | Roche | Cat# 11867423001, RRID:AB_390919 | WB (1:1000); IFA (1:1000). Only used in Figure 2. |
| Antibody | Mouse monoclonal clone TP3 anti-Toxoplasma | Abcam | Cat# ab8313, RRID:AB_306466 | WB (1:1000). |
| Antibody | Mouse monoclonal anti-GFP | Roche | Cat# 11814460001, RRID:AB_390913 | WB (1:1000). |
| Antibody | Rabbit polyclonal anti-mCherry | Abcam | Cat# ab167453, RRID:AB_2571870 | WB (1:1000). |
| Antibody | Rabbit monoclonal anti-SAG1 | PMID:3183382 | Provided by John Boothroyd Lab; WB (1:10,000). Only used in Figure 2. | |
| Antibody | Rabbit monoclonal clone 51-8 anti-thiophosphate ester | Abcam | Cat# ab92570, RRID:AB_10562142 | WB (1:5000). |
| Antibody | Mouse monoclonal clone 12G10 anti-tubulin | Developmental Studies Hybridoma Bank at the University of Iowa | RRID:AB_1157911 | Provided by Marc-Jan Gubbels Lab. WB (1:2000). |
| Antibody | Guinea pig monoclonal anti-CDPK1 | Covance | Custom antibody | WB (1/50,000). |
| Antibody | Rabbit monoclonal (C29F4) anti-HA | Cell Signaling Technology | Cat# 3724, RRID:AB_1549585 | WB (1:1000); IFA (1:1600). |
| Antibody | Mouse monoclonal clone 6D10 anti-MIC2 | PMID:10799515 | Provided by L. David Sibley Lab. WB (1:5000); IFA (1:2000). | |
| Antibody | Mouse anti-ROP1 | PMID:12467986 | Provided by Peter Bradley Lab. IFA (1:2000). | |
| Antibody | Mouse polyclonal anti-SAG1 | PMID:3183382 | Provided by L. David Sibley Lab. IFA (1:500). Used for invasion assays. | |
| Antibody | Rabbit polyclonal anti-GAP45 | Lampire Biological Laboratory | Provided by R. Drew Etheridge Lab. IFA (1:1000). Used for invasion assays. | |
| Antibody | Mouse monoclonal (16B12) anti-HA | BioLegend | Cat# 901533, RRID:AB_2565005 | WB (1:1000). |
| Antibody | Rabbit polyclonal clone WU1614 anti-ALD | PMID:16923803 | Provided by L. David Sibley Lab. WB (1:10,000). | |
| Antibody | Rabbit polyclonal anti-GAP45 | PMID:18312842 | Provided by Dominique Soldati-Favre Lab. WB (1:5000); IFA (1:5000). | |
| Antibody | Peroxidase-AffiniPure polyclonal Goat Anti-Rabbit IgG (H+L) | Jackson ImmunoResearch Laboratories | Cat# 111-035-003, RRID:AB_2313567 | WB (1/5000). |
| Antibody | Alexa Fluor 594 polyclonal Goat Anti-Rabbit IgG (H+L) | Life Technologies | IFA (1:1000). | |
| Antibody | Alexa Fluor 488 polyclonal Goat Anti-Mouse IgG (H+L) | Life Technologies | IFA (1:1000). | |
| Antibody | Alexa Fluor 488 polyclonal Goat Anti-Rabbit IgG (H+L) | Life Technologies | IFA (1:1000). | |
| Antibody | Alexa Fluor 594 polyclonal Goat Anti-Mouse IgG (H+L) | Life Technologies | IFA (1:1000). | |
| Antibody | IRDye 800CW Donkey anti-Guinea Pig IgG Secondary Antibody | LI-COR | LI-COR: 926-32411 | WB (1:10,000). |
| Antibody | IRDye 680RD Donkey anti-Guinea Pig IgG Secondary Antibody | LI-COR | LI-COR: 926-68077 | WB (1:10,000). |
| Antibody | IRDye 800CW Goat anti-Mouse IgG1-Specific Secondary Antibody | LI-COR | LI-COR: 926-32350 | WB (1:10,000). |
| Antibody | IRDye 680LT Goat anti-Mouse IgG Secondary Antibody | LI-COR | LI-COR: 926-68020 | WB (1:10,000). |
| Antibody | IRDye 800CW Goat anti-Rabbit IgG Secondary Antibody | LI-COR | LI-COR: 926-32211 | WB (1:10,000). |
| Antibody | IRDye 680LT Goat anti-Rabbit IgG Secondary Antibody | LI-COR | LI-COR: 926-68021 | WB (1:10,000). |
| Antibody | Alpaca monoclonal (1B7) anti-CDPK1 | PMID:26305940 | n/a | |
| Peptide, recombinant protein | IRDye 680RD Streptavidin | LI-COR | LI-COR: 926-68079 | WB (1:3000). |
| Peptide, recombinant protein | Aerolysin | PMID:26584919 | n/a | |
| Peptide, recombinant protein | Sequencing Grade Modified Trypsin | Promega | Promega: V5113 | |
| Chemical compound, drug | 3-Indoleacetic acid (auxin) | Sigma-Aldrich | Sigma-Aldrich: I2886-5G | |
| Chemical compound, drug | Compound 1 | PMID:12455981 | n/a | |
| Chemical compound, drug | Hoechst 33258 | Santa Cruz | Santa Cruz: sc-394039 | Egress assay (1:4000). |
| Chemical compound, drug | Hoechst 33342 | Invitrogen | Invitrogen: H3570 | IFA (1:20,000). |
| Chemical compound, drug | DAPI (4',6-diamidino-2-phenylindole, dihydrochloride) | Invitrogen | Invitrogen: D1306 | |
| Chemical compound, drug | Prolong Diamond | Thermo Fisher | Thermo Fisher: P36965 | |
| Chemical compound, drug | Zaprinast | Calbiochem | Calbiochem: 684500 | |
| Chemical compound, drug | A23187 | Calbiochem | Calbiochem: 100105 | |
| Chemical compound, drug | Myristic acid | Tokyo Chemical Industry | Cat# M0476 | |
| Chemical compound, drug | Alkyne-myristic acid (YnMyr) | Iris Biotech | Cat# RL-2055 | |
| Chemical compound, drug | Trypsin cleavable capture reagent | PMID:25807930; PMID:32618271 | This reagent was first reported as RTB in PMID:25807930. | |
| Chemical compound, drug | L-Proline for SILAC | Thermo Fisher Scientific | Thermo Fisher Scientific: 88211 | |
| Chemical compound, drug | Fetal bovine serum, dialyzed, US origin, One Shot format | Thermo Fisher Scientific | Thermo Fisher Scientific: A3382001 | |
| Chemical compound, drug | DMEM for SILAC | Thermo Fisher Scientific | Thermo Fisher Scientific: 88364 | |
| Chemical compound, drug | L-Arginine-HCl for SILAC | Thermo Fisher Scientific | Thermo Fisher Scientific: 89989 | |
| Chemical compound, drug | L-Lysine-2HCl for SILAC | Thermo Fisher Scientific | Thermo Fisher Scientific: 89987 | |
| Chemical compound, drug | L-Arginine-HCl, 13C6, 15N4 for SILAC | Thermo Fisher Scientific | Thermo Fisher Scientific: 89990 | |
| Chemical compound, drug | L-Lysine-2HCl, 13C6, 15N2 for SILAC | Thermo Fisher Scientific | Thermo Fisher Scientific: 88209 | |
| Chemical compound, drug | p-Nitrobenzyl mesylate (PNBM) | Epitomics | Epitomics # 3700-1 | |
| Chemical compound, drug | OXONE, monopersulfate compound | Sigma-Aldrich | Sigma-Aldrich: 228036-5G | |
| Chemical compound, drug | N6-furfuryladenosine (kinetin)–5′-O-[3- thiotriphosphate] sodium salt (KTPγS) | Axxora | Axxora: BLG-F008-05 | |
| Chemical compound, drug | Adenosine 5′-triphosphate disodium salt hydrate (ATP) | Sigma-Aldrich | Sigma-Aldrich: A6419-1G | |
| Chemical compound, drug | Guanosine 5ʹ-Triphosphate, Disodium Salt (GTP) | Calbiochem | Millipore: 371701 | |
| Chemical compound, drug | β-Casein from bovine milk | Sigma-Aldrich | Sigma-Aldrich: C6905 | |
| Chemical compound, drug | Biotin | Sigma-Aldrich | Sigma-Aldrich: B4501-1G | |
| Chemical compound, drug | PP1 Analog III, 3-MB-PP1 | Calbiochem | Sigma-Aldrich: 529582 | |
| Chemical compound, drug | Formaldehyde | Sigma-Aldrich | Sigma-Aldrich: F8775 | |
| Chemical compound, drug | Acrylamide | Sigma-Aldrich | Sigma-Aldrich: A4058 | |
| Chemical compound, drug | Sodium acrylate | AK Scientific | AK Scientific: R624 | |
| Chemical compound, drug | N,N'-Methylenebisacrylamide (BIS) | Sigma-Aldrich | Sigma-Aldrich: M1533 | |
| Chemical compound, drug | Tetramethyle thylenediamine (TEMED) |
Sigma-Aldrich | Sigma-Aldrich: T9281 | |
| Chemical compound, drug | Ammonium persulfate (APS) | Sigma-Aldrich | Sigma-Aldrich: A3678 | |
| Commercial assay or kit | S-trap micro | Protifi | Protifi: C02-micro-80 | |
| Commercial assay or kit | Pierce Quantitative Fluorometric Peptide Assay | Thermo Fisher Scientific | Thermo Fisher Scientific: 23290 | |
| Commercial assay or kit | TMTpro 16plex Label Reagent Set | Thermo Fisher Scientific | Thermo Fisher Scientific: A44522 | |
| Commercial assay or kit | EasyPep MS Sample Prep Kits - Maxi | Thermo Fisher Scientific | Thermo Fisher Scientific: A45734 | |
| Commercial assay or kit | High-Select TiO2 Phosphopeptide Enrichment Kit | Thermo Fisher Scientific | Thermo Fisher Scientific: A32993 | |
| Commercial assay or kit | High-Select Fe-NTA Phosphopeptide Enrichment Kit | Thermo Fisher Scientific | Thermo Fisher Scientific: A32992 | |
| Commercial assay or kit | Pierce High pH Reversed-Phase Peptide Fractionation Kit | Thermo Fisher Scientific | Thermo Fisher Scientific: 84868 | |
| Commercial assay or kit | Dynabeads MyOne Streptavidin C1 | Invitrogen | Cat# 65001 | |
| Commercial assay or kit | Bio-Rad DC assay | Bio-Rad | Bio-Rad: 5000116 | |
| Commercial assay or kit | Sep-Pak C18 Plus Short Cartridge, 360 mg Sorbent per Cartridge, 55–105 µm | Waters | Waters: WAT020515 | |
| Commercial assay or kit | SulfoLink Coupling Resin | Thermo Fisher Scientific | Thermo Fisher Scientific: 20401 | |
| Commercial assay or kit | Radiance Plus Chemiluminescent Substrate | Azure Biosystems | VWR: 10147-298 | |
| Commercial assay or kit | Pierce Anti-HA Magnetic Beads | Thermo Scientific | Thermo Scientific: 88836 | |
| Commercial assay or kit | Pierce Streptavidin Magnetic Beads | Thermo Scientific | Thermo Scientific: 88817 | |
| Recombinant DNA reagent | All plasmids used in this study are listed in Supplementary file 10 | This paper | ||
| Sequence-based reagent | All primers and oligonucleotides used in this study are listed in Supplementary file 10 | This paper | ||
| Software, algorithm | Proteome Discoverer 4.2 | Thermo Fisher | ||
| Software, algorithm | MaxQuant (version 1.5.0.25 and 1.5.2.8) | PMID:19029910 | RRID:SCR_014485 | Free software for searching of mass spectrometry acquisition files. |
| Software, algorithm | Perseus (version 1.5.0.9) | PMID:27348712 | RRID:SCR_015753 | Free software for processing of MaxQuant output files. |
| Software, algorithm | Scaffold DIA | Proteome Software | ||
| Software, algorithm | R version 4.0 | R Foundation for Statistical Computing | ||
| Software, algorithm | Prism 8 | GraphPad | ||
| Software, algorithm | Fiji | PMID:22743772 | ||
| Software, algorithm | ilastik | PMID:31570887 | ilastik.org | |
| Software, algorithm | HHPRED | PMID:29258817 | ||
| Software, algorithm | SnapGene | Dotmatics | https://www.snapgene.com/ | |
| Software, algorithm | ToxoDB | PMID:18003657 | ToxoDB.org | |
| Software, algorithm | Imaris (release 9.9) | Oxford Instruments | https://imaris.oxinst.com/ | |
| Other | DMEM, high glucose (media) | Life Technologies | Life Technologies: 11965118 | Materials and methods: Cell culture |
| Other | HALT protease inhibitor | Thermo Fisher | Thermo Fisher: 87786 | Materials and methods: Immunoprecipitation, lysis buffer |
| Other | HALT protease and phosphatase inhibitor | Thermo Fisher | Thermo Fisher: PI78440 | Materials and methods: Sub-minute resolution phosphoproteomics, lysis buffer |
| Other | Benzonase | Sigma-Aldrich | Sigma-Aldrich: E1014 | Materials and methods: Sub-minute resolution phosphoproteomics, lysis buffer |