Skip to content
Licensed Unlicensed Requires Authentication Published by De Gruyter August 8, 2005

Digestive versus regulatory proteases: on calpain action in vivo

  • Peter Friedrich and Zoltán Bozóky
From the journal Biological Chemistry


Calpains, the cytoplasmic Ca2+-activated regulatory proteases, have no simple and clearly definable cleavage site specificity, which is in sharp contrast to digestive (e.g., pancreatic) proteases. For calpains, an approximate 10-aa segment having a variety of sequences and spanning the scissile bond, governs proteolytic cleavage. This permissivity is a precondition for calpains to act on several different substrate proteins in the cell. The specificity of calpain action may be ensured by anchoring/targeting proteins. Intriguingly, the established endogenous inhibitor protein, calpastatin, might also serve as a storage site. Furthermore, specificity may be encoded in the ‘goodness’ of the undecapeptide sequence in substrate proteins. Novel approaches are needed to reveal how calpains find their substrates in cells at the proper time and location.


Corresponding author


Alexa, A., Bozoky, Z., Farkas, A., Tompa, P., and Friedrich, P. (2004). Contribution of distinct structural elements to activation of calpain by Ca2+ ions.J. Biol. Chem.279, 20118–20126.10.1074/jbc.M311969200Search in Google Scholar PubMed

Alto, N.M., Soderling, S.H., Hoshi, N., Langeberg, L.K., Fayos, R., Jennings, P.A., and Scott, J.D. (2003). Bioinformatic design of A-kinase anchoring protein-in silico: a potent and selective peptide antagonist of type II protein kinase A anchoring. Proc. Natl. Acad. Sci. USA100, 4445–4450.10.1073/pnas.0330734100Search in Google Scholar PubMed PubMed Central

Banik, N.L., Chou, C.H., Deibler, G.E., Krutzch, H.C., and Hogan, E.L. (1994). Peptide bond specificity of calpain: proteolysis of human myelin basic protein. J. Neurosci. Res.37, 489–496.10.1002/jnr.490370408Search in Google Scholar PubMed

Blomgren, K., Hallin, U., Andersson, A.L., Puka-Sundvall, M., Bahr, B.A., McRae, A., Saido, T.C., Kawashima, S., and Hagberg, H. (1999). Calpastatin is up-regulated in response to hypoxia and is a suicide substrate to calpain after neonatal cerebral hypoxia-ischemia. J. Biol. Chem.274, 14046–14052.10.1074/jbc.274.20.14046Search in Google Scholar PubMed

Coolican, S.A. and Hathaway, D.R. (1984). Effect of l-α-phosphatidylinositol on a vascular smooth muscle Ca2+-dependent protease. Reduction of the Ca2+ requirement for autolysis. J. Biol. Chem.259, 11627–11630.Search in Google Scholar

Dunker, A.K., Brown, C.J., Lawson, J.D., Iakoucheva, L.M., and Obradovic, Z. (2002). Intrinsic disorder and protein function. Biochemistry41, 6573–6582.10.1021/bi012159+Search in Google Scholar PubMed

Elce, J.S., Hegadorn, C. and Arthur, J.S. (1997). Autolysis, Ca2+ requirement, and heterodimer stability in m-calpain. J. Biol. Chem.272, 11268–11275.10.1074/jbc.272.17.11268Search in Google Scholar PubMed

Farkas, A., Tompa, P., Schad, E., Sinka, R., Jekely, G., and Friedrich, P. (2004). Autolytic activation and localization in Schneider cells (S2) of calpain B from Drosophila. Biochem. J.378, 299–305.10.1042/bj20031310Search in Google Scholar

Fernandez-Montalvan, A., Assfalg-Machleidt, I., Pfeiler, D., Fritz, H., Jochum, M., and Machleidt, W. (2004). Electrostatic interactions of domain III stabilize the inactive conformation of μ-calpain. Biochem. J.382, 607–617.10.1042/BJ20040731Search in Google Scholar PubMed PubMed Central

Friedrich, P. (2004). The intriguing Ca2+ requirement of calpain activation. Biochem. Biophys. Res. Commun.323, 1131–1133.10.1016/j.bbrc.2004.08.194Search in Google Scholar PubMed

Friedrich, P., Tompa, P., and Farkas, A. (2004). The calpain-system of Drosophila melanogaster: coming of age. Bioessays26, 1088–1096.10.1002/bies.20106Search in Google Scholar PubMed

Goll, D.E., Thompson, V.F., Li, H., Wei, W., and Cong, J. (2003). The calpain system. Physiol. Rev.83, 731–801.10.1152/physrev.00029.2002Search in Google Scholar

Hao, L.Y., Kameyama, A., Kuroki, S., Takano, J., Takano, E., Maki, M., and Kameyama, M. (2000). Calpastatin domain L is involved in the regulation of L-type Ca2+ channels in guinea pig cardiac myocytes. Biochem. Biophys. Res. Commun.279, 756–761.10.1006/bbrc.2000.4040Search in Google Scholar

Hosfield, C.M., Elce, J.S., Davies, P.L., and Jia, Z. (1999). Crystal structure of calpain reveals the structural basis for Ca2+-dependent protease activity and a novel mode of enzyme activation. EMBO J.18, 6880–6889.10.1093/emboj/18.24.6880Search in Google Scholar

Hosfield, C.M., Moldoveanu, T., Davies, P.L., Elce, J.S., and Jia, Z. (2001). Calpain mutants with increased Ca2+ sensitivity and implications for the role of the C2-like domain. J. Biol. Chem.276, 7404–7407.10.1074/jbc.M007352200Search in Google Scholar

Hosfield, C.M., Elce, J.S., and Jia, Z. (2004). Activation of calpain by Ca2+: roles of the large subunit N-terminal and domain III-IV linker peptides. J. Mol. Biol.343, 1049–1053.10.1016/j.jmb.2004.08.073Search in Google Scholar

Konno, T., Tanaka, N., Kataoka, M., Takano, E., and Maki, M. (1997). A circular dichroism study of preferential hydration and alcohol effects on a denatured protein, pig calpastatin domain I. Biochim. Biophys. Acta1342, 73–82.10.1016/S0167-4838(97)00092-7Search in Google Scholar

Ma, H., Yang, H.Q., Takano, E., Hatanaka, M., and Maki, M. (1994). Amino-terminal conserved region in proteinase inhibitor domain of calpastatin potentiates its calpain inhibitory activity by interacting with calmodulin-like domain of the proteinase. J. Biol. Chem.269, 24430–24436.10.1016/S0021-9258(19)51102-4Search in Google Scholar

Moldoveanu, T., Hosfield, C.M., Lim, D., Elce, J.S., Jia, Z., and Davies, P.L. (2002). A Ca2+ switch aligns the active site of calpain. Cell108, 649–660.10.1016/S0092-8674(02)00659-1Search in Google Scholar

Moldoveanu, T., Jia, Z., and Davies, P.L. (2004). Calpain activation by cooperative Ca2+ binding at two non-EF-hand sites. J. Biol. Chem.279, 6106–6114.10.1074/jbc.M310460200Search in Google Scholar PubMed

Mucsi, Z., Hudecz, F., Hollosi, M., Tompa, P., and Friedrich, P. (2003). Binding-induced folding transitions in calpastatin subdomains A and C. Protein Sci.12, 2327–2336.10.1110/ps.03138803Search in Google Scholar PubMed PubMed Central

Rawlings, N.D., Morton, F.R., Barret, A.J., and Bateman, A. (2005). MEROPS, the peptidase database. in Google Scholar

Reverter, D., Sorimachi, H., and Bode, W. (2001). The structure of calcium-free human m-calpain: implications for calcium activation and function. Trends Cardiovasc. Med.11, 222–229.10.1016/S1050-1738(01)00112-8Search in Google Scholar

Rosenmund, C., Carr, D.W., Bergeson, S.E., Nilaver, G., Scott, J.D., and Westbrook, G.L. (1994). Anchoring of protein kinase A is required for modulation of AMPA/kainate receptors on hippocampal neurons. Nature368, 853–856.10.1038/368853a0Search in Google Scholar

Sakai, K., Akanuma, H., Imahori, K., and Kawashima, S. (1987). A unique specificity of a calcium activated neutral protease indicated in histone hydrolysis. J. Biochem. (Tokyo)101, 911–918.10.1093/oxfordjournals.jbchem.a121959Search in Google Scholar

Sasaki, T., Kikuchi, T., Yumoto, N., Yoshimura, N., and Murachi, T. (1984). Comparative specificity and kinetic studies on porcine calpain I and calpain II with naturally occurring peptides and synthetic fluorogenic substrates. J. Biol. Chem.259, 12489–12494.10.1016/S0021-9258(18)90773-8Search in Google Scholar

Stabach, P.R., Cianci, C.D., Glantz, S.B., Zhang, Z., and Morrow, J.S. (1997). Site-directed mutagenesis of αII spectrin at codon 1175 modulates its μ-calpain susceptibility. Biochemistry36, 57–65.10.1021/bi962034iSearch in Google Scholar

Strobl, S., Fernandez-Catalan, C., Braun, M., Huber, R., Masumoto, H., Nakagawa, K., Irie, A., Sorimachi, H., Bourenkow, G., Bartunik, H., Suzuki, K., and Bode, W. (2000). The crystal structure of calcium-free human m-calpain suggests an electrostatic switch mechanism for activation by calcium. Proc. Natl. Acad. Sci. USA97, 588–592.10.1073/pnas.97.2.588Search in Google Scholar

Tompa, P. (2002). Intrinsically unstructured proteins. Trends Biochem. Sci.27, 527–533.10.1016/S0968-0004(02)02169-2Search in Google Scholar

Tompa, P., Emori, Y., Sorimachi, H., Suzuki, K., and Friedrich, P. (2001). Domain III of calpain is a Ca2+-regulated phospholipid-binding domain. Biochem. Biophys. Res. Commun.280, 1333–1339.10.1006/bbrc.2001.4279Search in Google Scholar PubMed

Tompa, P., Mucsi, Z., Orosz, G., and Friedrich, P. (2002). Calpastatin subdomains A and C are activators of calpain. J. Biol. Chem.277, 9022–9026.10.1074/jbc.C100700200Search in Google Scholar PubMed

Tompa, P., Buzder-Lantos, P., Tantos, A., Farkas, A., Szilagyi, A., Banoczi, Z., Hudecz, F., and Friedrich, P. (2004). On the sequential determinants of calpain cleavage. J. Biol. Chem.279, 20775–20785.10.1074/jbc.M313873200Search in Google Scholar PubMed

Wendt, A., Thompson, V.F., and Goll, D.E. (2004). Interaction of calpastatin with calpain: a review. Biol. Chem.385, 465–472.10.1515/BC.2004.054Search in Google Scholar PubMed

Wong, W. and Scott, J.D. (2004). AKAP signalling complexes: focal points in space and time. Nat. Rev. Mol. Cell Biol.5, 959–970.10.1038/nrm1527Search in Google Scholar PubMed

Zhang, W., Lane, R.D., and Mellgren, R.L. (1996). The major calpain isozymes are long-lived proteins. Design of an antisense strategy for calpain depletion in cultured cells. J. Biol. Chem.271, 18825–18830.10.1074/jbc.271.31.18825Search in Google Scholar PubMed

Published Online: 2005-08-08
Published in Print: 2005-07-01

©2005 by Walter de Gruyter Berlin New York

Downloaded on 27.2.2024 from
Scroll to top button