Androgen and glucocorticoid receptors interact with insulin degrading enzyme.
We recently identified a protein, designated receptor accessory factor (RAF), that interacts directly with and enhances the DNA binding of androgen (AR) and glucocorticoid (GR) receptor peptide fragments. To determine its identity, RAF was purified from HeLa cell extracts by anion-exchange and DNA affinity chromatography. RAF activity co-purified with a 110-kDa protein, the partial amino acid sequence of which shares 97.5% identity with insulin degrading enzyme (IDE), a metalloendoprotease implicated in the intracellular degradation of insulin. The identity of RAF was confirmed in gel shift assays by demonstrating that AR.RAF and GR.RAF DNA complexes shifted to a slower mobility in the presence of an IDE antibody. Purified preparations of RAF had insulin degrading activity, and this activity was inhibited by AR. In addition, the interaction of AR or GR with RAF was competed by insulin and bacitracin, a competitive inhibitor of IDE. The interactions of AR and GR with IDE may have important implications for both insulin- and steroid-mediated signaling.
Kupfer
S R
SR
Laboratories for Reproductive Biology, University of North Carolina School of Medicine, Chapel Hill 27599.
Wilson
E M
EM
French
F S
FS
D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007328:Insulin / Q000378:metabolism; D008666:Metalloendopeptidases; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000302:isolation & purification / Q000378:metabolism; D009838:Oligodeoxyribonucleotides; D011944:Receptors, Androgen / Q000378:metabolism*; D011965:Receptors, Glucocorticoid / Q000378:metabolism*; D015398:Signal Transduction
D007328:Insulin; D009687:Nuclear Proteins; D009838:Oligodeoxyribonucleotides; D011944:Receptors, Androgen; D011965:Receptors, Glucocorticoid; C072701:nuclear accessory protein, monkey; D010450:Endopeptidases; D008666:Metalloendopeptidases; C046583:pitrilysin