Abstract
A major area of calpain research involves the development of techniques to assess calpain activation in cells. Knowing physiologic conditions that allow the various calpain isoforms in cells to become activated should provide valuable information about their roles in cell functions. Several approaches have been tried, only a few of which will be discussed in this brief introduction.
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References
Rosser B. G., Powers S. P., and Gores G. J. (1993) Calpain activity increases in hepatocytes following addition of ATP. J. Biol. Chem. 268, 23,593–23,600.
Edelstein C. L., Wieder E. D., Yaqoob M. M., Gengaro P. E., Burke T. J., Nemenoff R. A., and Schrier R. W. (1995) The role of cysteine proteases in hypoxia-induced rat renal proximal tubular injury. Proc. Natl. Acad. Sci., U.S.A. 92, 7662–7666.
Saido T. C., Nagao S., Shiramine M., Tsukaguchi M., Sorimachi H., Murofushi H., Tsuchiya T., Ito H., and Suzuki K. (1992) Autolytic transition of mu-calpain upon activation as resolved by antibodies distinguishing between the pre-and post-autolysis forms. J. Biochem. (Tokyo) 111, 81–86.
Croall D. E., Slaughter C. A., Wortham H. S., Skelly C. M., DeOgny L., and Moomaw C. R. (1992) Polyclonal antisera specific for the proenzyme form of each calpain. Biochim. Biophys. Acta 1121, 47–53.
Schoenwaelder S. M., Kulkarni S., Salem H. H., Imajoh-Ohmi S., Yamao-Harigaya W., Saido T. C., and Jackson S. P. (1997) Distinct substrate specificities and functional roles for the 78-and 76-kDa forms of mu-calpain in human platelets. J. Biol. Chem. 272, 24,876–24,884.
Zhang W., Lane R. D., and Mellgren R. L. (1996) The major calpain isozymes are long-lived proteins. J. Biol. Chem. 271, 18,825–18,830.
Molinari M., Anagli J., and Carafoli E. (1994) Ca2+-activated neutral protease is active in the erythrocyte membrane in its nonautolyzed 80-kDa form. J. Biol. Chem. 269, 27,992–27,995.
Anagli J., Hagmann J., and Shaw E. (1991) Investigation of the role of calpain as a stimulus-response mediator in human platelets using new synthetic inhibitors. Biochem. J. 274, 497–502.
Anagli J., Hagmann J., and Shaw E. (1993) Affinity labelling of the Ca2+-acti-vated neutral proteinase (calpain) in intact human platelets. Biochem. J. 289, 93–99.
Mellgren R. L. and Lane R. D. (1988) Myocardial calpain 2 is inhibited by monoclonal antibodies specific for the small, noncatalytic subunit. Biochim. Biophys. Acta 954, 154–160.
Zhang W. and Mellgren R. L. (1996) Calpain subunits remain associated during catalysis. Biochem. Biophys. Res. Commun., 227, 890–896.
Mellgren R. L., Repetti A., Muck T. C., and Easly J. (1982) Rabbit skeletal muscle calcium-dependent protease requiring millimolar Ca2+. J. Biol. Chem. 257, 7203–7209.
Zhang W., Lu Q., Xie Z.-J., and Mellgren R. L. (1996) Inhibition of the growth of WI-38 fibroblasts by benzyloxycarbonyl-Leu-Leu-Tyr diazomethyl ketone: evidence that cleavage of p53 by a calpain-like protease is necessary for G1 to S-phase transition. Oncogene 14, 255–263.
Mellgren R. L. (1997) Evidence for participation of a calpain-like cysteine protease in cell cycle progression through late G1 phase. Biochem. Biophys. Res. Commun. 236, 555–558.
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© 2000 Humana Press Inc.
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Mellgren, R.L. (2000). A Radioimmunologic Technique for Assessing Calpain Activation in Cells. In: Elce, J.S. (eds) Calpain Methods and Protocols. Methods in Molecular Biology™, vol 144. Humana Press. https://doi.org/10.1385/1-59259-050-0:161
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DOI: https://doi.org/10.1385/1-59259-050-0:161
Publisher Name: Humana Press
Print ISBN: 978-0-89603-632-1
Online ISBN: 978-1-59259-050-6
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