Identification, purification, and characterization of a PA700-dependent activator of the proteasome

George N. DeMartino, Rita J. Proske, Carolyn R. Moomaw, Anita A. Strong, Xiaoling Song, Hiroshi Hisamatsu, Keiji Tanaka, Clive A. Slaughter

Research output: Contribution to journalArticle

124 Citations (Scopus)

Abstract

The activity of the intracellular protease, the proteasome, is modulated by a number of specific regulatory proteins. One such regulator, PA700, is a 700,000-Da multisubunit protein that activates hydrolytic activities of the proteasome via a mechanism that involves the ATP-dependent formation of a proteasome-PA700 complex. Four subunits of PA700 have been shown previously to be members of a protein family that contains a consensus sequence for ATP binding, and purified PA700 expresses ATPase activity. We report here the identification, purification, and initial characterization of a new modulator of the proteasome. The modulator has no direct effect on the activity of the proteasome, but enhances PA700 activation of the proteasome by up to 8-fold. This activation is associated with the formation of a proteasome/PA700- containing complex that is significantly larger than that formed in its absence. The modulator has a native M(r) of ~300,000, as determined by gel filtration chromatography, and is composed of three electrophoretically distinct subunits with M(r) values of 50,000, 42,000, and 27,000 (p50, p42, and p27, respectively). Amino acid sequence analysis of the subunits shows that p50 and p42 are members of the same ATP-binding protein family found in PA700. The p50 subunit is identical to TBP1, a protein previously reported to interact with human immunodeficiency virus Tat protein (Nelbock, P., Dillion, P. J., Perkins, A., and Rosen, C. A. (1990) Science 248, 1650-1653), while the p42 subunit seems to be a new member of the family. The p27 subunit has no significant sequence similarity to any previously described protein. Both p50 and p42, but not p27, were also identified as components of PA700, increasing the number of ATP-binding protein family members in this complex to six. Thus, p50 and p42 are subunits common to two protein complexes that regulate the proteasome. The PA700-dependent proteasome activator represents a new member of a growing list of proteins that regulate proteasome activity.

Original languageEnglish (US)
Pages (from-to)3112-3118
Number of pages7
JournalJournal of Biological Chemistry
Volume271
Issue number6
DOIs
StatePublished - Feb 9 1996
Externally publishedYes

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Proteasome Endopeptidase Complex
Purification
Adenosine Triphosphate
Proteins
Modulators
Carrier Proteins
tat Gene Products
Chemical activation
Human Immunodeficiency Virus Proteins
PA700 proteasome activator
Protein Sequence Analysis
Consensus Sequence
Chromatography
Gel Chromatography
Viruses
Adenosine Triphosphatases
Peptide Hydrolases
Gels
Amino Acids

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Identification, purification, and characterization of a PA700-dependent activator of the proteasome. / DeMartino, George N.; Proske, Rita J.; Moomaw, Carolyn R.; Strong, Anita A.; Song, Xiaoling; Hisamatsu, Hiroshi; Tanaka, Keiji; Slaughter, Clive A.

In: Journal of Biological Chemistry, Vol. 271, No. 6, 09.02.1996, p. 3112-3118.

Research output: Contribution to journalArticle

DeMartino, GN, Proske, RJ, Moomaw, CR, Strong, AA, Song, X, Hisamatsu, H, Tanaka, K & Slaughter, CA 1996, 'Identification, purification, and characterization of a PA700-dependent activator of the proteasome', Journal of Biological Chemistry, vol. 271, no. 6, pp. 3112-3118. https://doi.org/10.1074/jbc.271.6.3112
DeMartino, George N. ; Proske, Rita J. ; Moomaw, Carolyn R. ; Strong, Anita A. ; Song, Xiaoling ; Hisamatsu, Hiroshi ; Tanaka, Keiji ; Slaughter, Clive A. / Identification, purification, and characterization of a PA700-dependent activator of the proteasome. In: Journal of Biological Chemistry. 1996 ; Vol. 271, No. 6. pp. 3112-3118.
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AU - Strong, Anita A.

AU - Song, Xiaoling

AU - Hisamatsu, Hiroshi

AU - Tanaka, Keiji

AU - Slaughter, Clive A.

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