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A proteomic strategy for the identification of SGT‐interacting proteins in prostate cancer cells (539.6)
Author(s) -
Prieto Luis
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.539.6
Subject(s) - lncap , tetratricopeptide , chemistry , heat shock protein , nuclear receptor , microbiology and biotechnology , hsp90 , receptor , signal transducing adaptor protein , cancer cell , biology , biochemistry , gene , cancer , transcription factor , genetics
A Proteomic Strategy for the Identification of SGTAα‐ Interacting Proteins in Prostate Cancer Cells Molecular chaperones facilitate proper folding and regulation of steroid hormone receptors (SHRs). Upon proper folding, SHRs bind its ligand with high affinity, translocate to the nucleus and initiate gene expression. A fairly novel cochaperone, human small glutamine rich TPR (tetratricopeptide repeat) containing protein alpha (SGTA), is a down‐regulator of aAndrogen (AR), gGlucocorticoid (GR), and pProgesterone (PR) receptors. It binds to heat shock protein (Hsp) 70 and 90 kDa. Additionally, SGTA plays a role in cellular processes such as cell cycle progression and apoptosis. Therefore, a top‐down proteomic approach was used to determine novel interactors in LNCaP human prostate cancer cells line LNCaP. Recombinant FLAG‐A tagged ‐SGTA version was purified using a FLAG peptide and further analyzed for unknown interacting proteins with liquid chromatography mass spectrometry (LC‐MS/MS). A comparison of normal (150 mM) versus high (300 mM) salt concentration was used to determine strong interactors. Preliminary studies have identified RanBP2 E3 SUMO‐Ligase and microtubule to be present along with SGTA in both concentrations. Thus, future studies aim to validate these interactions with SGTA using nickel purification and assess their functional cellular relevance, which will contribute to the understanding of the role SGTA plays within the chaperoning pathway.