Identification of phosphotyrosine (pTyr)-containing T cell proteins
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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 (ncbitaxon:9606)
SUBMITTER:
Amnon Altman
PROVIDER: MSV000083562 | MassIVE | Wed Mar 13 11:19:00 GMT 2019
REPOSITORIES: MassIVE
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