title: Ubiquitinated proteasome inhibitor is a component of the 26 S proteasome complex.
abstract: Western blot analysis, using a polyclonal antibody to the 240-kDa endogenous inhibitor of the 20 S proteasome, revealed that the inhibitor is a component of the 26 S complex. Although isolated inhibitor displayed a single 40-kDa band on SDS-PAGE, the antibody detected a 55-kDa component in the 26 S proteasome complex. Ubiquitin polyclonal antibody recognized the same 55-kDa component but did not react with free 40-kDa inhibitor subunit. Addition of purified 40-kDa inhibitor to a ubiquitin ligating system also generated the 55-kDa species. In crude erythrocyte extracts, most of the inhibitor migrated at 55 kDa in the presence of ATP but shifted to 40 kDa in the absence of ATP, consistent with removal of ubiquitin. It is suggested that ubiquitination of the inhibitor may be involved in regulating assembly and/or activity of the 26 S proteasome complex.
authors.affiliation: Department of Cell Biology and Anatomy, New York Medical College, Valhalla, New York 10595.
authors.forename: X S
authors.initials: XS
authors.lastname: Li
authors.forename: J D
authors.initials: JD
authors.lastname: Etlinger
mesh_terms: D015153:Blotting, Western; D003546:Cysteine Endopeptidases / Q000737:chemistry*; D015853:Cysteine Proteinase Inhibitors / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000201:enzymology; D006801:Humans; D009097:Multienzyme Complexes / Q000737:chemistry*; D046988:Proteasome Endopeptidase Complex; D016622:Silver Staining; D014452:Ubiquitins / Q000737:chemistry*
chemical_list: D015853:Cysteine Proteinase Inhibitors; D009097:Multienzyme Complexes; D014452:Ubiquitins; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex