Proteasome-mediated degradation of tau proteins occurs independently of the chymotrypsin-like activity by a nonprocessive pathway.
20S proteasomes form the proteolytic core of the 26S proteasome responsible for degradation of substrates of the ubiquitin-proteasome pathway. In addition, 20S proteasomes have themselves been linked to degradation of intracellular proteins. This multienzyme complex expresses three distinct catalytic sites, each with unique substrate specificity. The contribution of these sites to overall proteolysis remains unclear. Also unclear is the kinetic mechanism of degradation. Studies with denatured or covalently modified proteins suggest that degradation is nonprocessive in some cases and processive in others. We sought greater insight into these questions by analyzing degradation of tau proteins and beta-casein. Tau proteins were readily degraded by bovine pituitary proteasomes. Degradation yielded large quantities of intermediates, which were more abundant as tau concentration was increased, indicating that degradation occurred by a nonprocessive pathway. Similar findings were observed for degradation of beta-casein. Experiments with inhibitors demonstrated that degradation of both full-length tau and the intermediates derived from it was largely dependent on the trypsin-like activity. A combination of inhibitors against the trypsin-like and glutamyl activities almost completely blocked tau degradation, while inhibitors active toward the chymotrypsin-like activity had minimal effects on degradation of tau and intermediates derived from it. These findings are discussed with respect to the contribution of the three catalytic sites to overall intracellular proteolysis, the factors contributing to nonprocessive degradation, and the implications of this type of pathway for intracellular proteolysis.
Cardozo
Christopher
C
Department of Medicine, Mount Sinai School of Medicine, 1 Gustave L. Levy Place, New York, NY 10029, USA. chris.cardozo@mssm.edu
Michaud
Charlene
C
D000818:Animals; D020134:Catalytic Domain; D002417:Cattle; D002918:Chymotrypsin / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D009097:Multienzyme Complexes / Q000302:isolation & purification / Q000378:metabolism*; D010902:Pituitary Gland / Q000201:enzymology; D046988:Proteasome Endopeptidase Complex; D020033:Protein Isoforms / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D013154:Spleen / Q000201:enzymology; D014357:Trypsin / Q000378:metabolism*; D025801:Ubiquitin / Q000378:metabolism; D016875:tau Proteins / Q000378:metabolism*
D009097:Multienzyme Complexes; D020033:Protein Isoforms; D011994:Recombinant Proteins; D025801:Ubiquitin; D016875:tau Proteins; D002918:Chymotrypsin; D014357:Trypsin; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex