1. AR is transnitrosylated by HSP90.
AR is transnitrosylated by HSP90. A, HSP90β forms a complex with AR and eNOS. LNCaP cells were lysed and subjected to immunoprecipitation with anti-HSP90β or isotype control [immunoglobulin G (IgG)] antibodies. Protein complexes were immunoblotted with the indicated antibodies. B and C, purified HSP90β (B) or AR (C) protein was exposed to CysNO (100 μmol/L) and resultant S-nitrosylated protein was desalted and then mixed with un-nitrosylated purified protein counterpart. Biotin switch assay was performed to detect the protein transnitrosylation. Input amounts of AR and HSP90β were verified in each reaction. D and E, transnitrosylation of AR in situ. PC3 cells were transfected with pcDNA3-Flag-AR alone (D) or together with pCMV-Myc-HSP90β (E). Cells were exposed to CysNO (200 μmol/L), and cell lysates were subjected to the biotin switch assay.
Yu Qin et al. Cancer Res 2013;73:
©2013 by American Association for Cancer Research