Altered proteasome structure, function, and oxidation in aged muscle.
The proteasome is the main protease for degrading oxidized proteins. We asked whether altered proteasome function contributes to the accumulation of oxidized muscle proteins with aging. Proteasome structure, function, and oxidation state were compared in young and aged F344BN rat fast-twitch skeletal muscle. In proteasome-enriched homogenates from aged muscle, we observed a two- to threefold increase in content of the 20S proteasome that was due to a corresponding increase in immunoproteasome. Content of the regulatory proteins, PA700 and PA28, relative to the 20S were reduced 75% with aging. Upon addition of exogenous PA700, there was a twofold increase in peptide hydrolysis in aged muscle, suggesting the endogenous content of PA700 is inadequate for complete activation of the 20S. Measures of catalytic activity showed a 50% reduction in specific activity for proteasome-enriched homogenates with aging. With purification of the 20S, proteasome specific activity was equivalent between ages, indicating that endogenous regulators inhibit proteasome in aged muscle. Significantly less degradation of oxidized calmodulin by the 20S from aged muscle was observed. Partial rescue of activity for aged 20S by DTT implies oxidation of functionally significant cysteines. These results demonstrate significant age-related changes in proteasome structure, function, and oxidation state that could inhibit removal of oxidized proteins.
Ferrington
Deborah A
DA
Department of Ophthalmology, 380 Lions Research Bldg., 2001 6th Street SE, University of Minnesota, Minneapolis, MN 55455, USA. ferri013@umn.edu
Husom
Aimee D
AD
Thompson
LaDora V
LV
D000375:Aging; D000818:Animals; D002147:Calmodulin / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D002417:Cattle; D004229:Dithiothreitol / Q000494:pharmacology; D006868:Hydrolysis; D008297:Male; D009124:Muscle Proteins / Q000737:chemistry* / Q000502:physiology*; D018482:Muscle, Skeletal / Q000737:chemistry*; D010084:Oxidation-Reduction; D046988:Proteasome Endopeptidase Complex / Q000008:administration & dosage / Q000032:analysis / Q000737:chemistry* / Q000502:physiology*; D051381:Rats; D011916:Rats, Inbred F344
D002147:Calmodulin; D009124:Muscle Proteins; C506159:PA28 protein, rat; C085834:PA700 proteasome activator; D046988:Proteasome Endopeptidase Complex; D004229:Dithiothreitol