Studies on mechanisms of calpain activation in the cell

2004 Fiscal Year Final Research Report Summary

• Project/Area Number: 14380309
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Functional biochemistry
• Research Institution: Tokyo Metropolitan Institute of Gerontology
• Principal Investigator: INOMATA Mitsushi Tokyo Metropolitan Foundation for Research on Aging and Promotion of Human Welfare, Tokyo Metropolitan Institute of Gerontology, 福祉振興財団・東京都老人総合研究所, 研究員 (30142649)
• Co-Investigator(Kenkyū-buntansha): SORIMACHI Hiroyuki The Tokyo Metropolitan Institute of Medical Science (Rinshoken), Tokyo Metropolitan Organization for Medical Research, Department of Enzymatic Regulation for Cell Functions, Department Head, 東京都臨床医学総合研究所, 副参事研究員 (10211327)
• Project Period (FY): 2002 – 2004
• Keywords: calpain / fodrin / talin / calpastatin / calcium / cyteskeltal protein / modulator protease / GFP

Research Abstract

Calpain is a "modulator protease" involved in Ca^<2+>-mediated signal transduction processes essential for cellular functions. Physiological functions, although yet unclear, of calpains are so important that a deficient calpain activity causes various diseases including muscular dystrophies, ischemia diseases, cancers, diabetes, and Alzheimer's disease. In these diseases, inhibition of calpain activity is often reported to improve the pathological states of patients, thus indicating the importance of calpain inhibitors and understanding of mechanisms of calpain activation. In this study, we expressed recombinant proteins corresponding to the protease domain of calpain and to EGFP-tagged C105S dominant negative m-calpain, and used to study molecular mechanisms of the regulation of calpain activity. To our surprise, the protease domain of m-calpain showed Ca^<2+>-dependent protease activity, indicating that the protease domain itself binds Ca^<2+>. This is a very important information as to how calpain activity is regulated. Moreover, using C105S-dominanto negative m-calpain, proteolysis of cyteskeletal proteins such as fodrin and talin showed differential susceptibility to m- and μ-calpains. Calpastatin, which is the highly specific and powerful endogenous inhibitor protein for calpain, did not suppress proteolysis of these proteins. These results strongly suggest that for proteolysis of calpain and its regulation, membrane, rather than calpastatin, is very important factor.

Research Products

• [Journal Article] Involvement of calpain in osteoclastic bone resorption (2005)
  • Author(s): Hayashi, M., Koshihara, Y., Ishibashi, H., Yamamoto, S., Tsubuki, S., Saido, TC., Kawashima, S., Inomata, M.
  • Journal Title: J.Biochem 137 Pages: 331-338
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Inhibitory NK receptor Ly49Q is expressed on subsets of dendritic cells in a cellular maturation-and cytokine stimulation-dependent manner. (2005)
  • Author(s): Toyama-Sorimachi, N., Omatsu, Y., Onoda, A., Tsujimura, Y., Iyoda, T., Maki, A., Sorimachi, H., Dohi, T., Taki, S., Inaba, K., Karasuyama, H.
  • Journal Title: J.Immun. 174 Pages: 4621-4629
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Involvement of calpain in osteoclastic bone resorption. (2005)
  • Author(s): Hayashi, M., Koshihara, Y., Ishibashi, H., Yamamoto.S., Tsubuki, S., Saido, T.C., Kawashima, S., Inomata, M.
  • Journal Title: J.Biochem. 137 Pages: 331-338
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Possible functions of p94 in connectin-mediated signaling pathways in skeletal muscle cells (2005)
  • Author(s): Ojima, K., Ono, Y., Hata, S., Koyama, S., Doi, N., Sorimachi, H.
  • Journal Title: J.Muscle Res.Cell Motil (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Calpain. (2004)
  • Author(s): Sorimachi, H., Ono, Y.
  • Journal Title: Encyclopedia of Biological Chemistry(eds., W.J.Lennarz, M.D.Lane)(Elsevier, Oxford) 1 Pages: 300-306
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Degradation of fodrin by m-calpain in fibroblasts adhering to fibrillar collagen I gel. (2004)
  • Author(s): Sato, K., Hattori, S., Irie, S., Sorimachi, H., Inomata, M., Kawashima, S.
  • Journal Title: J.Biochem. 136 Pages: 777-785
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Neoculin as a New Taste-modifying Protein Occurring in the Fruit of Curculigo latifolia. (2004)
  • Author(s): Shirasuka, Y., Nakajima, K., Asakura, T., Yamashita, H., Yamamoto, A., Hata, S., Nagata, S., Abo, M., Sorimachi, H., Abe, K.
  • Journal Title: Biosci.Biotechnol.Biochem. 68 Pages: 1403-1407
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Induction and myofibrillar targeting of CARP, and suppression of the Nkx2.5 pathway in the MDM mouse with impaired titin-based signaling. (2004)
  • Author(s): Witt, C.C.^*, Ono, Y.^*, Puschmann, E., McNabb, M., Wu, Y., Gotthardt, M., Witt, S.H., Haak, M., Labeit, D., Gregorio, C.C., Sorimachi, H., Granzier, H., Labeit, S.
  • Journal Title: J.Mol.Biol. 336 Pages: 145-154
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Possible regulation of the conventional calpain system by skeletal muscle-specific calpain, p94/calpain3. (2004)
  • Author(s): Ono, Y., Kakinuma, K., Torii, F., Irie, A., Nakagawa, K., Labeit, S., Abe, K., Suzuki, K., Sorimachi, H.
  • Journal Title: J.Biol.Chem. 279 Pages: 2761-2771
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Calpain. (2004)
  • Author(s): Sorimachi, H., Ono, Y.
  • Journal Title: Encyclopedia of Biological Chemistry(eds., W.J.Lennarz, M.D.Lane)(Elsevier, Oxford) vol.1 Pages: 300-306
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Induction and myofibrillar targeting of CARP, and suppression of the Nkx2.5 pathway in the MDM mouse with impaired titin-based signaling. (2004)
  • Author(s): Witt, C.C.^*, Ono, Y.^*, Puschmann, E., McNabb, M., Wu, Y., Gotthardt, M., Witt, S.H., Haak, M., Labeit, D., Gregorio, C.C., Sorimachi, H., Granzier, H., Labeit, S.(^*equally contributed first authors)
  • Journal Title: J.Mol.Biol. 336 Pages: 145-154
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Possible regulation of the conventional calpain system by skeletal muscle-specific calpain, p94/calpain 3. (2004)
  • Author(s): Ono, Y., Kakinuma, K., Torii, F., Irie, A., Nakagawa, K., Labeit, S., Abe, K., Suzuki, K., Sorimachi, H.
  • Journal Title: J.Biol.Chem. 279 Pages: 2761-2771
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Structure, activation, and biology of calpain. (2004)
  • Author(s): Suzuki, K., Hata, S., Kawabata, Y., Sorimachi, H.
  • Journal Title: Diabetes 53 Pages: S12-S18
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Newly identified exons encoding novel variants of p94/calpain3 are expressed ubiquitously and overlap the α-glucosidase C gene. (2003)
  • Author(s): Kawabata, Y., Hata, S., Ono, Y., Ito, Y., Suzuki, K., Abe, K., Sorimachi, H.
  • Journal Title: FEBS Lett. 555 Pages: 623-630
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Heterogeneous nuclear ribonucleoprotein K interacts with and is proteolyzed by calpain in vivo. (2003)
  • Author(s): Kimura, E., Abe, K., Suzuki, K., Sorimachi, H.
  • Journal Title: Biosci.Biotechnol.Biochem. 67 Pages: 3786-1796
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Flexibility analysis and structure comparison of two crystal forms of calcium-free human m-calpain. (2002)
  • Author(s): Reverter, D., Braun, M., Femandez-Catalan, C., Strobl, S., Sorimachi, H., Bode, W.
  • Journal Title: Biol Chem. 383 Pages: 1415-1422
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] C/EBP_is required for proteolytic cleavage of cyclin A by calpain 3 in myeloid precursor cells. (2002)
  • Author(s): Welm, A., L, Timchenko, N.A., Ono, Y., Sorimachi, H., Radomska, H.S., Tenen, D.G., Lekstrom-Himes, J., Darlington, G.J.
  • Journal Title: J.Biol.Chem. 277 Pages: 33848-33856
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Defects of non-lysosomal proteolysis : Calpain3 deficiency. (2002)
  • Author(s): Sorimachi, H., Beckmann, J.S.
  • Journal Title: Structural and Molecular Basis of Skeletal Muscle Diseases(ed., Karpati.G.) (ISN Neuropath.Press.Basel) Pages: 148-153
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Stable expression of calpain 3 from a muscle transgene in vivo : Immature muscle in transgenic mice suggests a role for calpain 3 in muscle maturation. (2002)
  • Author(s): Spencer, M.J., Guyon, J.R., Sorimachi, H., Potts, A., Richard, I., Herasse, M., Chamberlain, J., Dalkilic, I., Kunkel, L.M., Beckmann, J.S.
  • Journal Title: Proc.Natl.Acad.Sci.U.S.A. 929 Pages: 8874-8879
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Muscle-specific RING finger 1 interacts with titin to regulate sarcomeric M-line and thick filaments structure and may have nuclear functions via its interaction with glucocorticoid (2002)
  • Author(s): McElhinny, A.S., Kakinuma, K., Sorimachi, H., Labeit, S., Gregorio, C.C.
  • Journal Title: J.Cell Biol. 157 Pages: 125-136
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Comparison of Lp82- and m-calpain-mediated proteolysis during cataractogenesis in Shumiya cataract rat (SCR). (2002)
  • Author(s): Inomata, M., Hayashi, M., Ito, Y., Matsubara, Y., Takehana, M., Kawashima, S., Shumiya, S.
  • Journal Title: Curr.Eye Res. 25 Pages: 207-213
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Klotho protein deficiency leads to overactivation of μ-calpain. (2002)
  • Author(s): Manya, H., Inomata, M., Fujimori, T., Domae, N., Sato, Y., Takio, K., Mabeshima, Y., Endo, T.
  • Journal Title: J.Biol.Chem. 277 Pages: 35503-35508
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Flexibility analysis and structure comparison of two crystal forms of calcium-free human m-calpain. (2002)
  • Author(s): Reverter, D., Braun, M., Fernandez-Catalan, C., Strobl, S., Sorimuchi, H., Bode, W.
  • Journal Title: Biol Chem. 383 Pages: 1415-1422
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] C/EBP_is required for proteolytic cleavage of cyclin A by calpain 3 in myeloid precursor cells. (2002)
  • Author(s): Welm, A.L., Timchenko, N.A., Ono, Y., Sorimachi, H., Radomska, H.S., Tenen, D.G., Lekstrom-Himes, J., Darlington, G.J.
  • Journal Title: J.Biol.Chem. 277 Pages: 33848-33856
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Defects of non-lysosomal proteolysis : Calpain3 deficiency. (2002)
  • Author(s): Sorimachi, H., Beckmann, J.S.
  • Journal Title: Structural and Molecular Basis of Skeletal Muscle Diseases(ed, Karpati, G.)(ISN Neuropath.Press, Basel) Pages: 148-153
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Stable expression of calpain 3 from a muscle transgene in vivo : Immature muscle in transgenic mice suggests a role for calpain 3 in muscle maturation. (2002)
  • Author(s): Spencer, M.J., Guyon, J.R., Sorimachi, H., Potts, A., Richard, I., Herasse, M., Chamberlain, J., Dalkilic, I., Kunkel, L.M., Beckmann, J.S.
  • Journal Title: Proc.Natl.Acad.Sci.U.S.A. 99 Pages: 8874-8879
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Muscle-specific RING finger 1 interacts with titin to regulate sarcomeric M-line and thick filament structure and may have nuclear functions via its interaction with glucocorticoid modulatory element binding protein 1. (2002)
  • Author(s): McElhinny, A.S., Kakinuma, K., Sorimachi, H., Labeit, S., Gregorio, C.C.
  • Journal Title: J.Cell Biol. 157 Pages: 125-136
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Comparison of Lp82- and m-calpain-mediated proteolysis during cataractogenesis in Shumiya cataract rat(SCR). (2002)
  • Author(s): Inomata, M., Hayashi, M., Ito, Y., Matsubara, Y., Takehana, M., Kawashima, S., Shumiya, S.
  • Journal Title: Curr.Eye Res. 25 Pages: 207-213
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Klotho protein deficiency leads to overactivation of mu-calpain. (2002)
  • Author(s): Manya, H., Inomata, M., Fujimori, T., Dohmae, N., Sato, Y., Takio, K., Nabeshima, Y., Endo, T.
  • Journal Title: J.Biol.Chem. 277 Pages: 35503-35508
  • Description: 「研究成果報告書概要(欧文)」より