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Phosphoproteomic quantitation and causal analysis reveal pathways in GPVI/ITAM-mediated platelet activation programs.
Babur, Özgün; Melrose, Alexander R; Cunliffe, Jennifer M; Klimek, John; Pang, Jiaqing; Sepp, Anna-Liisa I; Zilberman-Rudenko, Jevgenia; Tassi Yunga, Samuel; Zheng, Tony; Parra-Izquierdo, Iván; Minnier, Jessica; McCarty, Owen J T; Demir, Emek; Reddy, Ashok P; Wilmarth, Phillip A; David, Larry L; Aslan, Joseph E.
Afiliación
  • Babur Ö; Department of Molecular and Medical Genetics.
  • Melrose AR; Computational Biology Program.
  • Cunliffe JM; Knight Cardiovascular Institute.
  • Klimek J; Proteomics Shared Resource.
  • Pang J; Proteomics Shared Resource.
  • Sepp AI; Department of Biomedical Engineering.
  • Zilberman-Rudenko J; Department of Biomedical Engineering.
  • Tassi Yunga S; Department of Biomedical Engineering.
  • Zheng T; Cancer Early Detection & Advanced Research Center.
  • Parra-Izquierdo I; Department of Biomedical Engineering.
  • Minnier J; Knight Cardiovascular Institute.
  • McCarty OJT; Department of Biomedical Engineering.
  • Demir E; Knight Cancer Institute, and.
  • Reddy AP; Department of Biomedical Engineering.
  • Wilmarth PA; Department of Molecular and Medical Genetics.
  • David LL; Computational Biology Program.
  • Aslan JE; Proteomics Shared Resource.
Blood ; 136(20): 2346-2358, 2020 11 12.
Article en En | MEDLINE | ID: mdl-32640021
ABSTRACT
Platelets engage cues of pending vascular injury through coordinated adhesion, secretion, and aggregation responses. These rapid, progressive changes in platelet form and function are orchestrated downstream of specific receptors on the platelet surface and through intracellular signaling mechanisms that remain systematically undefined. This study brings together cell physiological and phosphoproteomics methods to profile signaling mechanisms downstream of the immunotyrosine activation motif (ITAM) platelet collagen receptor GPVI. Peptide tandem mass tag (TMT) labeling, sample multiplexing, synchronous precursor selection (SPS), and triple stage tandem mass spectrometry (MS3) detected >3000 significant (false discovery rate < 0.05) phosphorylation events on >1300 proteins over conditions initiating and progressing GPVI-mediated platelet activation. With literature-guided causal inference tools, >300 site-specific signaling relations were mapped from phosphoproteomics data among key and emerging GPVI effectors (ie, FcRγ, Syk, PLCγ2, PKCδ, DAPP1). Through signaling validation studies and functional screening, other less-characterized targets were also considered within the context of GPVI/ITAM pathways, including Ras/MAPK axis proteins (ie, KSR1, SOS1, STAT1, Hsp27). Highly regulated GPVI/ITAM targets out of context of curated knowledge were also illuminated, including a system of >40 Rab GTPases and associated regulatory proteins, where GPVI-mediated Rab7 S72 phosphorylation and endolysosomal maturation were blocked by TAK1 inhibition. In addition to serving as a model for generating and testing hypotheses from omics datasets, this study puts forth a means to identify hemostatic effectors, biomarkers, and therapeutic targets relevant to thrombosis, vascular inflammation, and other platelet-associated disease states.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Algoritmos / Glicoproteínas de Membrana Plaquetaria / Activación Plaquetaria / Proteómica Límite: Animals / Humans Idioma: En Revista: Blood Año: 2020 Tipo del documento: Article

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Algoritmos / Glicoproteínas de Membrana Plaquetaria / Activación Plaquetaria / Proteómica Límite: Animals / Humans Idioma: En Revista: Blood Año: 2020 Tipo del documento: Article
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