Conformational Remodeling of Proteasomal Substrates by PA700, the 19 S Regulatory Complex of the 26 S Proteasome

PA700, the 19 S regulatory complex of the 26 S proteasome, plays a central role in the recognition and efficient degradation of misfolded proteins. PA700 promotes degradation by recruiting proteasomal substrates utilizing polyubiquitin chains and chaperone-like binding activities and by opening the...

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Bibliographic Details
Published in:	The Journal of biological chemistry Vol. 277; no. 30; pp. 26815 - 26820
Main Authors:	Liu, Chang-wei, Millen, Linda, Roman, Tracie B., Xiong, Hai, Gilbert, Hiram F., Noiva, Robert, DeMartino, George N., Thomas, Philip J.
Format:	Journal Article
Language:	English
Published:	United States Elsevier Inc 26-07-2002 American Society for Biochemistry and Molecular Biology
Subjects:	Adenosine Triphosphatases - chemistry Animals Catalytic Domain Cattle Chymotrypsin - pharmacology Disulfides - chemistry Dose-Response Relationship, Drug Endopeptidases - chemistry Enzyme Inhibitors - pharmacology Ligands Methotrexate - pharmacology Oxygen - metabolism Peptides - chemistry Proteasome Endopeptidase Complex Protein Conformation Protein Folding Ribonuclease, Pancreatic - chemistry Time Factors
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Summary:	PA700, the 19 S regulatory complex of the 26 S proteasome, plays a central role in the recognition and efficient degradation of misfolded proteins. PA700 promotes degradation by recruiting proteasomal substrates utilizing polyubiquitin chains and chaperone-like binding activities and by opening the access to the core of the 20 S proteasome to promote degradation. Here we provide evidence that PA700 in addition to binding misfolded protein substrates also acts to remodel their conformation prior to proteolysis. Scrambled RNase A (scRNase A), a misfolded protein, only slowly refolds spontaneously into an active form because of the rate-limiting unfolding of misfolded disulfide isomers. Notably, PA700 accelerates the rate of reactivation of scRNase A, consistent with its ability to increase the exposure of these disulfide bonds to the solvent. In this regard, PA700 also exposes otherwise buried sites to digestion by exogenous chymotrypsin in a polyubiquitinated enzymatically active substrate, pentaubiquitinated dihydrofolate reductase, Ub5DHFR. The dihydrofolate reductase ligand methotrexate counters the ability of PA700 to promote digestion by chymotrypsin. Together, these results indicate that in addition to increasing substrate affinity and opening the access channel to the catalytic sites, PA700 activates proteasomal degradation by remodeling the conformation of protein substrates.
ISSN:	0021-9258 1083-351X
DOI:	10.1074/jbc.M201782200