Elsevier

Methods in Enzymology

Volume 244, 1994, Pages 331-350
Methods in Enzymology

[24] Multicatalytic endopeptidase complex: Proteasome

https://doi.org/10.1016/0076-6879(94)44026-3Get rights and content

Publisher Summary

This chapter describes assay, purification, and properties of proteasome. The multicatalytic endopeptidase complex or proteasome is a 700-kDa multisubunit enzyme complex that is widely distributed in eukaryotic cells. The multicatalytic endopeptidase complex, either by itself or as the catalytic core of the 26S proteinase complex, is assumed to play an important role in ubiquitin-dependent and ubiquitin independent nonlysosomal pathways of protein turnover, including the degradation of regulatory proteins and the processing of antigens for presentation by the major histocompatibility complex (MHC) class I pathway. Different substrates have been used to assay the endopeptidase activities of the complex purified from many different sources. Its substrates include proteins, peptides, and synthetic peptides and also the enzyme is active over a range of neutral to weakly alkaline pH values in a variety of different buffers. The 7-amino-4-methylcoumarin leaving group provides the most sensitive assay and these synthetic peptides are often used at concentrations below their Km values, which are in the range of 0.1-1 mM.

References (55)

  • A.J. Rivett

    Arch. Biochem. Biophys.

    (1989)
  • M. Rechsteiner et al.

    J. Biol. Chem.

    (1993)
  • A.J. Rivett

    J. Biol. Chem.

    (1985)
  • K. Tanaka et al.

    J. Biol. Chem.

    (1988)
  • P.E. Falkenburg et al.

    J. Biol. Chem.

    (1989)
  • A.J. Rivett

    J. Biol. Chem.

    (1989)
  • A.J. Rivett

    Arch. Biochem. Biophys.

    (1985)
  • A.J. Rivett et al.

    Arch. Biochem. Biophys.

    (1990)
  • R.E. Pacifici et al.

    J. Biol. Chem.

    (1993)
  • K. Tanaka et al.

    J. Biol. Chem.

    (1986)
  • K.B. Hendil et al.

    J. Biochem. Biophys. Methods

    (1991)
  • M.J. McGuire et al.

    Biochim. Biophys. Acta.

    (1986)
  • L.W. Lee et al.

    Biochim. Biophys. Acta

    (1990)
  • M.E. Pereira et al.

    J. Biol. Chem.

    (1992)
  • M. Chu-Ping et al.

    Biochim. Biophys. Acta

    (1992)
  • M.M. Bradford

    Anal. Biochem.

    (1976)
  • L. Hoffman et al.

    J. Biol. Chem.

    (1992)
  • J. Driscoll et al.

    J. Biol. Chem.

    (1990)
  • A. Grziwa et al.

    FEBS Lett.

    (1991)
  • K.S. Lilley et al.

    FEBS Lett.

    (1990)
  • Y. Saitoh et al.

    Biochem. Biophys. Res. Commun.

    (1989)
  • W. Hilt et al.

    J. Biol. Chem.

    (1993)
  • W. Heinemeyer et al.

    J. Biol. Chem.

    (1993)
  • S. Jentsch

    Trends Cell Biol.

    (1992)
  • A.J. Rivett et al.

    Curr. Biol.

    (1993)
  • M. Rechsteiner

    Cell (Cambridge, Mass.)

    (1991)
  • P.E. Falkenberg et al.

    Nature (London)

    (1988)
  • Cited by (0)

    View full text