Appendix 1—key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Chemical compound, drug | AccuPrime Pfx master mix | Thermo Fisher | 12344040 | |
| Chemical compound, drug | Pfu Ultra High-Fidelity DNA Polymerase | Agilent | 600380 | |
| Chemical compound, drug | Fugene transfection reagent | Thermo Fisher | 10362100 | |
| Chemical compound, drug | SIGMAFAST Protease Inhibitor Cocktail Tablets, EDTA-Free | Sigma | P8830 | |
| Chemical compound, drug | Beta-mercaptoethanol | Sigma | M3148-100ML | |
| Chemical compound, drug | PMSF | Sigma | P7626 | |
| Chemical compound, drug | Guanosine 5′-diphosphate (GDP) sodium salt hydrate | Sigma | G7127-100MG | |
| Chemical compound, drug | Guanosine 5′-triphosphate (GTP) sodium salt hydrate | Sigma | G8877-250MG | |
| Chemical compound, drug | Sodium deoxycholate | Sigma | D6750 | |
| Chemical compound, drug | C12E10 (Polyoxyethylene (10) lauryl ether) | Sigma | P9769 | |
| Chemical compound, drug | CHAPS | Thermo Fisher | J6735909 | |
| Chemical compound, drug | 1,2-dioleoyl-sn-glycero-3-phosphocholine (18:1 DOPC) | Avanti | 850375 C | |
| Chemical compound, drug | 1,2-dioleoyl-sn-glycero-3-phospho-L-serine (18:1 DOPS) | Avanti | 840035 C | |
| Chemical compound, drug | 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (18:1 Δ9-Cis) DOPE | Avanti | 850725 C | |
| Chemical compound, drug | L-α-phosphatidylinositol-4,5-bisphosphate (Brain PI(4,5)P2) | Avanti | 840046 X | |
| Chemical compound, drug | phosphatidylinositol 4,5-bisphosphate 18:0/20:4 (PI(4,5)P2) | Echelon | P-4524 | |
| Chemical compound, drug | 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N- [4-(p-maleimidomethyl)cyclohexane-carboxamide] (18:1 MCC-PE) |
Avanti | 780201 C | |
| Chemical compound, drug | 10 x PBS [pH 7.4] | Corning | 46–013 CM | |
| Chemical compound, drug | Trolox | Cayman Chemicals | 10011659 | |
| Chemical compound, drug | Dyomics 647 maleimide | Dyomics | 647 P1-03 | |
| Chemical compound, drug | Cy3 maleimide | Cytiva | PA23031 | |
| Chemical compound, drug | Cy5 maleimide | Cytiva | PA15131 | |
| Chemical compound, drug | Cy3 Mono NHS Ester | Cytiva | PA13101 | |
| Chemical compound, drug | SNAP-Surface Alexa Fluor 488 | NEB | S9129S | |
| Chemical compound, drug | SNAP-Surface Alexa Fluor 546 | NEB | S9132S | |
| Chemical compound, drug | SNAP-Surface Alexa Fluor 647 | NEB | S9136S | |
| Chemical compound, drug | Coenzyme A | Sigma | C3019 | |
| Chemical compound, drug | Sulfuric acid | Sigma | 58105–2.5 L-PC | |
| Peptide, recombinant protein | glucose oxidase (Aspergillus niger, 225 U/mg) | Biophoretics | B01357.02 | |
| Peptide, recombinant protein | catalase (bovine liver) | Sigma | C40-100MG | |
| Peptide, recombinant protein | 10 mg/mL beta casein solution | ThermoFisher | 37528 | |
| Peptide, recombinant protein | LPETGG | ELIM Biopharm | custom peptide synthesis | >95% purity |
| Peptide, recombinant protein | CSDGG(pY)MDMSKDESID(pY) VPMLDMKGDIKYADIE |
ELIM Biopharm | custom peptide synthesis | >95% purity |
| Peptide, recombinant protein | CSDGG(pY)MDMSKDESID(pY) VPMLDMKGDIKYADIE-Alexa488 |
ELIM Biopharm | custom peptide synthesis | >95% purity |
| Sequence-based reagent |
CCTTTTTGGTAgcaGATGCG TCTACTAAAATGCATGGTG |
Integrated DNA Technologies (IDT) | FW_ ybbr-PIK3R1 (R358A) | |
| Sequence-based reagent |
GTAGACGCATCtgcTACCAAAAA GGTCCCGTCTGCTGTATC |
IDT | RV_ ybbr-PIK3R1 (R358A) | |
| Sequence-based reagent |
CTTTTCTTGTCgcgGAGAGC AGTAAACAGGGCTGC |
IDT | FW_ ybbr-PIK3R1 (R358A) | |
| Sequence-based reagent |
GTTTACTGCTCTCcgcGACAAG AAAAGTGCCATCTCGCTTC |
IDT | RV_ ybbr-PIK3R1 (R358A) | |
| Sequence-based reagent |
CAAGTCGAGGTGGAgatgacTTT CTTCCTGTATTGAAAGAAATCTTGG |
IDT | FW_ his6-TEV-PIK3CB(K532D,K533D) | |
| Sequence-based reagent |
CAATACAGGAAGAAAgtcatcTCCAC CTCGACTTGACACATTAGCAC |
IDT | RV_ his6-TEV-PIK3CB(K532D,K533D) | |
| Recombinant DNA reagent | his10-SUMO3-GGGGG-Rac1(1-192aa) | This paper | pSH752 | Bacterial protein expression plasmid |
| Recombinant DNA reagent | his6-MBP-N10-TEV-GGGGG-P-Rex1 (40-405aa, DH-PH) |
This paper | pSH658 | Bacterial protein expression plasmid |
| Recombinant DNA reagent | his10-SUMO3-p67/phox TRP (Rac1-GTP sensor) | This paper | pSH823 | Bacterial protein expression plasmid |
| Recombinant DNA reagent | his6-GST-TEV-nSH2 PIK3R1(322-440aa)-Cys | This paper | pSH615 | Bacterial protein expression plasmid |
| Recombinant DNA reagent | his6-SUMO-Btk(PH-TH,R49S/K52S)-SNAP | This paper | pSH1313 | Bacterial protein expression plasmid |
| Recombinant DNA reagent | his6-MBP-N10-TEV-GGGGG-Grp1 (261-387aa) | This paper | pSH558 | Bacterial protein expression plasmid |
| Recombinant DNA reagent | his6-TEV-SenP2 (368-589aa, SUMO protease) | This paper | pSH653 | Bacterial protein expression plasmid |
| Recombinant DNA reagent | his6-TEV-PIK3CB (1-1070aa) | This paper | pSH541 | Baculovirus expression plasmid |
| Recombinant DNA reagent | ybbr-PIK3R1 (1-724aa) | This paper | pSH743 | Baculovirus expression plasmid |
| recombinant DNA reagent | ybbr-PIK3R1 (FVLR->FVLA, R358A) | This paper | pSH1045 | Baculovirus expression plasmid |
| Recombinant DNA reagent | ybbr-PIK3R1 (FVLR->FVLA, R358A) | This paper | pSH1046 | Baculovirus expression plasmid |
| Recombinant DNA reagent | his6-TEV-PIK3CB (1-1070aa; K532D,K533D) | This paper | pSH1094 | Baculovirus expression plasmid |
| Recombinant DNA reagent | his6-Gg2, Gb1 (DUAL FastBac) | PMID:34452907 | pSH414 | Baculovirus expression plasmid |
| Recombinant DNA reagent | his6-Gg2, SNAP-Gb1 (DUAL FastBac) | PMID:34452907 | pSH651 | Baculovirus expression plasmid |
| Recombinant DNA reagent | PIK3CG(p110g), TwinStrept-his10-TEV-ybbr-PIK3R5(p101) | PMID:36842083 | HP29 | Baculovirus expression plasmid |
| Software, algorithm | GraphPad Prism 9 | GraphPad | https://www.graphpad.com | |
| Software, algorithm | Chimera | UCSF | https://www.rbvi.ucsf.edu/chimera/ | |
| Software, algorithm | ImageJ/Fiji | ImageJ | https://imagej.net/software/fiji/ | |
| Software, algorithm | Nikon NIS elements | Nikon | https://www.microscope.healthcare.nikon.com/products/software/nis-elements | |
| Cell line(Spodoptera frugiperda) | Sf9 insect cells | Expression Systems | 94–001 S | |
| Cell line (Trichoplusiani) | High five insect cells | UC Berkeley Barker Hall Tissue Culture Facility | High five insect cells | |
| Other | ESF 921 Serum-Free Insect Cell Culture media | Expression Systems | 96-001-01 | Media for insect cell culture |
| Other | Fetal Bovine serum | Seradigm | 1500–500 | Media for insect cell culture |
| Other | Hellmanex III cleaning solution | Fisher | 14-385-864 | Reagent for cleaning coverslips prior to Pirahna etching |