Proteomics

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Identification of phosphotyrosine (pTyr)-containing T cell proteins


ABSTRACT: This project aimed at identifying phosphotyrosine (pTyr)-containing T cell proteins that bind directly to the pTyr-binding pocket in the N-terminal C2 domain of protein kinase C-theta. We used an immobilized recombinant GST-C2 fusion protein to pull-down proteins present in lysates of stimulated human leukemic Jurkat T cell line. After eluting and separating C2-bound proteins by SDS-PAGE, protein bands in the ~60-70 kDa range detected by Coomassie blue staining or anti-pTr immunoblotting were excised from the gel, destained, and subjected to trypsinization. The resulting peptides were extracted, dried, dissolved in 0.1% formic acid, and analyzed by online nano flow LC-MS/MS.

INSTRUMENT(S): LTQ Orbitrap, 1200 series LC/MSD SL

ORGANISM(S): Homo sapiens  

SUBMITTER: Amnon Altman   

PROVIDER: MSV000083562 | Massive | Wed Mar 13 11:19:00 GMT 2019

REPOSITORIES: MassIVE

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