Recognition of misfolding proteins by PA700, the regulatory subcomplex of the 26 S proteasome

Elizabeth Strickland, Kevin Hakala, Philip J. Thomas, George N. DeMartino

Research output: Contribution to journalArticle

157 Citations (Scopus)

Abstract

The 26 S proteasome is a large protease complex that catalyzes the degradation of both native and misfolded proteins. These proteins are known to interact with PA700, the regulatory subcomplex of the 26 S proteasome, via a covalently attached polyubiquitin chain. Here we provide evidence for an additional ubiquitin-independent mode of substrate recognition by PA700. PA700 prevents the aggregation of three incompletely folded, nonubiquitinated substrates: the ΔF-508 mutant form of cystic fibrosis transmembrane regulator, nucleotide binding domain 1, insulin B chain, and citrate synthase. This function does not require ATP hydrolysis. The stoichiometry required for this function, the effect of PA700 on the lag phase of aggregation, and the temporal specificity of PA700 in this process all indicate that PA700 interacts with a subpopulation of non-native conformations that is either particularly aggregation-prone or nucleates misassociation reactions. The inhibition of off-pathway self-association reactions is also reflected in the ability of PA700 to promote refolding of citrate synthase. These results provide evidence that, in addition to binding polyubiquitin chains, PA700 contains a site(s) that recognizes and interacts with misfolded or partially denatured polypeptides. This feature supplies an additional level of substrate specificity to the 26 S proteasome and a means by which substrates are maintained in a soluble state until refolding or degradation is complete.

Original languageEnglish (US)
Pages (from-to)5565-5572
Number of pages8
JournalJournal of Biological Chemistry
Volume275
Issue number8
DOIs
StatePublished - Feb 25 2000

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Proteasome Endopeptidase Complex
Polyubiquitin
Citrate (si)-Synthase
Agglomeration
Substrates
Proteins
Ubiquitin
Substrate Specificity
Cystic Fibrosis
Degradation
Hydrolysis
Peptide Hydrolases
Nucleotides
Adenosine Triphosphate
Stoichiometry
Insulin
Conformations
Peptides
Association reactions

ASJC Scopus subject areas

  • Biochemistry

Cite this

Recognition of misfolding proteins by PA700, the regulatory subcomplex of the 26 S proteasome. / Strickland, Elizabeth; Hakala, Kevin; Thomas, Philip J.; DeMartino, George N.

In: Journal of Biological Chemistry, Vol. 275, No. 8, 25.02.2000, p. 5565-5572.

Research output: Contribution to journalArticle

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