Elsevier

Methods in Enzymology

Volume 244, 1994, Pages 685-700
Methods in Enzymology

[49] Cystatins

https://doi.org/10.1016/0076-6879(94)44051-4Get rights and content

Publisher Summary

This chapter highlights cystatins, which are functionally active cysteine peptidase inhibitor evolutionarily related to chicken cystatin. It focuses on human cystatins and also describes isolation and assay of cystatins in other mammals. Three types or families of cystatins have been recognized. Families 1 and 2 contain low molecular weight cystatins, which are proteins of about 100–120 amino acid residues. The proteins in family 1 are normally intracellular, being synthesized without signal peptides, and contain no disulfide bonds. In contrast, those in family 2 are synthesized with signal peptides, and are secreted proteins that contain disulfide bonds. The family 3 cystatins are the kininogens, much larger proteins that contain three cystatin domains, two of which are functional and unrelated domains. As all cystatins inhibit papain by formation of tight, reversible complexes, assays for cystatin activity are based on the inhibition of the peptidase activity of this enzyme, which results from preincubation with the inhibitor. The size difference between human family 1 cystatins and the family 2 cystatins can readily be detected by sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE) using a discontinuous system and separation gels containing 20% acrylamide.

References (50)

  • K. Abe et al.

    J. Biol. Chem.

    (1987)
  • J.F. Morrison

    Trends Biochem. Sci.

    (1982)
  • G. Mancini et al.

    Immunochemistry

    (1965)
  • M. Abrahamson et al.

    J. Biol. Chem.

    (1986)
  • J.P. Freije et al.

    J. Biol. Chem.

    (1993)
  • H. Schägger et al.

    Anal. Biochem.

    (1987)
  • M. Järvinen

    J. Invest. Dermatol.

    (1978)
  • M. Järvinen et al.

    Biochim. Biophys. Acta

    (1982)
  • B. Turk et al.

    J. Biol. Chem.

    (1993)
  • P.A. Shaw et al.

    J. Biol. Chem.

    (1988)
  • M. Solem et al.

    Biochem. Biophys. Res. Commun.

    (1990)
  • A. Esnard et al.

    FEBS Lett.

    (1988)
  • M. Hirado et al.

    FEBS Lett.

    (1985)
  • M.D. Poulik et al.

    Mol. Immunol.

    (1981)
  • M. Sasaki et al.

    Biochem. Biophys. Res. Commun.

    (1977)
  • W. Müller-Esterl et al.

    Trends Biochem. Sci.

    (1986)
  • J.T. Finkenstaedt
  • A. J. Barrett, this series, Vol. 80, p....
  • N.D. Rawlings et al.

    J. Mol. Evol.

    (1990)
  • A. Ritonja et al.

    Biochem. J.

    (1987)
  • H. Saito et al.

    Biochemistry

    (1989)
  • A.J. Barrett et al.
  • V. Turk et al.

    FEES Lett.

    (1992)
  • M.O. Funk et al.

    Int. J. Pept. Protein Res.

    (1979)
  • A.J. Barrett et al.

    Biochem. J.

    (1982)
  • Cited by (186)

    • Structure determinants defining the specificity of papain-like cysteine proteases

      2022, Computational and Structural Biotechnology Journal
    View all citing articles on Scopus
    View full text