Conformational remodeling of proteasomal substrates by PA700, the 19 S regulatory complex of the 26 S proteasome

Chang Wei Liu, Linda Millen, Tracie B. Roman, Hai Xiong, Hiram F. Gilbert, Robert Noiva, George N. Demartino, Philip J. Thomas

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)26815-26820
Number of pages6
JournalJournal of Biological Chemistry
Volume277
Issue number30
DOIs
StatePublished - Jul 26 2002

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Pancreatic Ribonuclease
Tetrahydrofolate Dehydrogenase
Chymotrypsin
Disulfides
Proteolysis
Digestion
Polyubiquitin
Protein Conformation
Proteasome Endopeptidase Complex
Substrates
Degradation
Methotrexate
Catalytic Domain
Carrier Proteins
Conformations
Ligands
Proteins
Isomers
ATP dependent 26S protease

ASJC Scopus subject areas

  • Biochemistry

Cite this

Conformational remodeling of proteasomal substrates by PA700, the 19 S regulatory complex of the 26 S proteasome. / Liu, Chang Wei; Millen, Linda; Roman, Tracie B.; Xiong, Hai; Gilbert, Hiram F.; Noiva, Robert; Demartino, George N.; Thomas, Philip J.

In: Journal of Biological Chemistry, Vol. 277, No. 30, 26.07.2002, p. 26815-26820.

Research output: Contribution to journalArticle

Liu, Chang Wei ; Millen, Linda ; Roman, Tracie B. ; Xiong, Hai ; Gilbert, Hiram F. ; Noiva, Robert ; Demartino, George N. ; Thomas, Philip J. / Conformational remodeling of proteasomal substrates by PA700, the 19 S regulatory complex of the 26 S proteasome. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 30. pp. 26815-26820.
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