Dataset Information

- Identification of Gαi2-interacting proteins via affinity pull-down and quantitative mass-spectrometry analysis.

ABSTRACT: In studying patients with activating GNAI2 mutations, we observed T cell hyperresponsiveness that was independent of cAMP suppression. Therefore, we hypothesized that Gαi2 utilizes alternative signal transduction pathways which might be delineated by identifying its interacting proteins in T cells. To accomplish this, we performed affinity pulldown followed by quantitative mass-spectrometry analysis. Using this approach, we identified both well-known and previously reported interactors of Gαi2 as well as 128 previously unreported interactors including RASA2. We validated RASA2, a GTPase-activating protein for RAS, and PP2A-A/C as Gαi2 effector targets by several methods: overexpression-coimmunoprecipitations using 293T cells, endogenous coimmunoprecipitations from patient T cells, and by mixing-pulldown experiments using purified recombinant proteins. Additional imaging experiments confirmed Gαi2-RASA2 interactions within living cells and delineated that activating mutations of Gαi2 sequester RASA2 toward the plasma membrane to promote RAS activation for T cell growth and proliferation.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human) Escherichia Coli

TISSUE(S): T Cell, Blood

SUBMITTER: David Anderson

LAB HEAD: Helen C. Su

PROVIDER: PXD048980 | Pride | 2024-09-20

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	Chicken.fasta	Fasta
	Human071717.fasta	Fasta
	UniprotEscherichiacolistrainK12.fasta	Fasta
	allSpectra.CID.ITMS.iso_0.apl	Other
	allSpectra.CID.ITMS.iso_1.apl	Other

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Publications

Germline mutations in a G protein identify signaling cross-talk in T cells.

Ham Hyoungjun H Jing Huie H Lamborn Ian T IT Kober Megan M MM Koval Alexey A Berchiche Yamina A YA Anderson D Eric DE Druey Kirk M KM Mandl Judith N JN Isidor Bertrand B Ferreira Carlos R CR Freeman Alexandra F AF Ganesan Sundar S Karsak Meliha M Mustillo Peter J PJ Teo Juliana J Zolkipli-Cunningham Zarazuela Z Chatron Nicolas N Lecoquierre François F Oler Andrew J AJ Schmid Jana Pachlopnik JP Kuhns Douglas B DB Xu Xuehua X Hauck Fabian F Al-Herz Waleed W Wagner Matias M Terhal Paulien A PA Muurinen Mari M Barlogis Vincent V Cruz Phillip P Danielson Jeffrey J Stewart Helen H Loid Petra P Rading Sebastian S Keren Boris B Pfundt Rolph R Zarember Kol A KA Vill Katharina K Potocki Lorraine L Olivier Kenneth N KN Lesca Gaetan G Faivre Laurence L Wong Melanie M Puel Anne A Chou Janet J Tusseau Maud M Moutsopoulos Niki M NM Matthews Helen F HF Simons Cas C Taft Ryan J RJ Soldatos Ariane A Masle-Farquhar Etienne E Pittaluga Stefania S Brink Robert R Fink Danielle L DL Kong Heidi H HH Kabat Juraj J Kim Woo Sung WS Bierhals Tatjana T Meguro Kazuyuki K Hsu Amy P AP Gu Jingwen J Stoddard Jennifer J Banos-Pinero Benito B Slack Maria M Trivellin Giampaolo G Mazel Benoît B Soomann Maarja M Li Samuel S Watts Val J VJ Stratakis Constantine A CA Rodriguez-Quevedo Maria F MF Bruel Ange-Line AL Lipsanen-Nyman Marita M Saultier Paul P Jain Rashmi R Lehalle Daphne D Torres Daniel D Sullivan Kathleen E KE Barbarot Sébastien S Neu Axel A Duffourd Yannis Y Similuk Morgan M McWalter Kirsty K Blanc Pierre P Bézieau Stéphane S Jin Tian T Geha Raif S RS Casanova Jean-Laurent JL Makitie Outi M OM Kubisch Christian C Edery Patrick P Christodoulou John J Germain Ronald N RN Goodnow Christopher C CC Sakmar Thomas P TP Billadeau Daniel D DD Küry Sébastien S Katanaev Vladimir L VL Zhang Yu Y Lenardo Michael J MJ Su Helen C HC

Science (New York, N.Y.) 20240920 6715

Humans with monogenic inborn errors responsible for extreme disease phenotypes can reveal essential physiological pathways. We investigated germline mutations in GNAI2, which encodes Gαi2, a key component in heterotrimeric G protein signal transduction usually thought to regulate adenylyl cyclase-mediated cyclic adenosine monophosphate (cAMP) production. Patients with activating Gαi2 mutations had clinical presentations that included impaired immunity. Mutant Gα ...[more]

PMID: 39298586

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