Involvement of the proteasome in various degradative processes in mammalian cells.
Eukaryotic cells contain a 700-kDa proteolytic complex (the "proteasome" or multicatalytic endopeptidase complex), whose role in intracellular protein breakdown is unclear. It has been suggested that the proteasome functions in the rapid degradation of oxidant-damaged proteins and in the ATP-dependent proteolytic pathway. To test these possibilities, oxidant-damaged hemoglobin and albumin were produced by treating hemoglobin and albumin with phenylhydrazine, with hydroxyl radicals, or with both hydroxyl and superoxide radicals. After oxidant damage, these proteins were degraded more rapidly in erythrocyte extracts and also by the purified proteasome. However, complete removal of proteasomes from these extracts by immunoprecipitation (or inhibitors of its proteolytic activity) did not reduce the breakdown of oxidant-damaged hemoglobin and decreased degradation of hydroxyl- and superoxide-treated proteins by only 30-40%. Thus, erythrocytes must contain another proteolytic system for degradation of oxidant-damaged proteins. In contrast, immunoprecipitation of proteasomes with polyclonal or monoclonal antibodies prevented the ATP/ubiquitin-dependent degradation of lysozyme and also blocked the ATP-stimulated degradation of ubiquitin-conjugated lysozyme in reticulocyte and skeletal muscle extracts. These data indicate a critical role of the proteasome in the degradation of ubiquitin-conjugated proteins and suggest that the proteasome is associated with or is a component of the larger ubiquitin-conjugate-degrading enzyme complex.
Matthews
W
W
Department of Cellular and Molecular Physiology, Harvard Medical School, Boston, MA 02115.
Driscoll
J
J
Tanaka
K
K
Ichihara
A
A
Goldberg
A L
AL
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D003546:Cysteine Endopeptidases / Q000502:physiology*; D004912:Erythrocytes / Q000201:enzymology*; D005609:Free Radicals; D006801:Humans; D066298:In Vitro Techniques; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000502:physiology*; D010084:Oxidation-Reduction; D011233:Precipitin Tests; D046988:Proteasome Endopeptidase Complex; D011506:Proteins / Q000378:metabolism*; D011817:Rabbits; D014452:Ubiquitins / Q000502:physiology
D000911:Antibodies, Monoclonal; D005609:Free Radicals; D009097:Multienzyme Complexes; D011506:Proteins; D014452:Ubiquitins; D000255:Adenosine Triphosphate; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex