Purification of PA700, the 19S regulatory complex of the 26S proteasome

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

The 26S proteasome is a 2,400,000-Da protease complex that selectively degrades proteins modified by polyubiquitin chains. The 26S proteasome is composed of two 700,000-Da multisubunit complexes: the 20S proteasome, which serves as the proteolytic core of the complex, and PA700, an ATPase regulatory complex responsible for the binding, modification, and delivery of substrates to the proteolytic chamber. Thus, PA700 mediates multiple functions essential for ubiquitin-dependent proteolysis by the 26S proteasome. This chapter reviews briefly the structure and function of PA700, details the methodology for its large-scale purification from mammalian tissues, and describes a simple functional PA700 assay based on the stimulation of proteasome activity.

Original languageEnglish (US)
Pages (from-to)295-306
Number of pages12
JournalMethods in Enzymology
Volume398
DOIs
StatePublished - 2005

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Purification
Proteasome Endopeptidase Complex
Polyubiquitin
Proteolysis
Ubiquitin
Adenosine Triphosphatases
Assays
Peptide Hydrolases
Tissue
Substrates
ATP dependent 26S protease
Proteins
PA700 proteasome activator

ASJC Scopus subject areas

  • Biochemistry

Cite this

Purification of PA700, the 19S regulatory complex of the 26S proteasome. / DeMartino, George N.

In: Methods in Enzymology, Vol. 398, 2005, p. 295-306.

Research output: Contribution to journalArticle

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