2007 Fiscal Year Final Research Report Summary
Functional analysis of digestive tract-specific calpains in terms of functional foods.
| Project/Area Number |
18380085
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Food science
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| Research Institution | Tokyo Metropolitan Organization for Medical Research |
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Principal Investigator |
SORIMACHI Hiroyuki Tokyo Metropolitan Organization for Medical Research, the Tokyo Metropolitan Institute of Medical Science, 副参事研究員 (10211327)
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| Project Period (FY) |
2006 – 2007
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| Keywords | calpain / enzyme / protein / proteolysis / mutant mouse / digestive tract / membrane traffiching / COPI |
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| Research Abstract |
Calpains are a cytosolic Ca^<2+>-regulated cysteine protease. Calpains directly interact with intracellular proteins to modify or transform functions and/or activities of their substrates, and are thus called "modulator protease". Mammals have 15 independent genes for calpains, about half of which are expressed tissue specifically. These tissue-specific calpain are expected to be involved in specific functions developed in the specific tissues, and, therefore, their physiological functions are of great interest. Two of the tissue specific calpains, nCL-2/-2' and nCL-4, are predominantly expressed in digestive tracts, especially, in the epithelial mucus secreting cells. These cells protect the stomach and intestines from acid/proteases and bacteria. Therefore, these calpains' functions are considered to be related with mucus secretion mechanisms.
To elucidate physiological functions of these calpains, we constructed nCL-2: C105S "knock in" mice, which express a protease inactive mutant of nCL-2 instead of wild type nCL-2 under endogenous expression controls. A missense mutation corresponding to the Cys105 to Ser was introduced in the mouse genomic DNA, and neomycin-resistance gene (neoR) flanked by loxP sequences was inserted in the vicinity of the C105S mutation. After homologously targeted mice were obtained, they were crossed with Cre-recombinase expressing transgenic mice to excise neoR, resulting in nCL-2: C105S knock-in mice.
Using these knock-in mice and their tissues, various biochemical, cell-biological, and molecular biological analyses have been performed. As a result, nCL-2 was shown to form not only monomer but also homo-oligomers via C2-domain like domain (domain III). This novel structure suggests unique functions for nCL-2 in digestive tracts.
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[Journal Article] Muscle RING-Finger Protein MuRF1 as a Connector of Muscle Energy Metabolism and Protein Synthesis.2008
Author(s)
Koyama, S.*, Hata, S.*, Witt, C. C.*, Ono, Y., Lerche, S., Ojima, K., Chiba, T., Doi, N., Kitamura, F., Tanaka, K., Abe, K., Witt, S., Rybin, V., Gasch, A., Franz, T., Labeit, S., and Sorimachi, H. (*equally contributed first authors)
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Journal Title
J. Mol. Biol. 376, (peer-reviewed)
Pages: 1224-1236
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Stomach-specific calpain, nCL-2/calpain 8, is active without calpain regulatory subunit, and oligomerizes through C2-like domains.2007
Author(s)
Hata, S., Doi, N., Kitamura, F., and Sorimachi, H.
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Journal Title
J. Biol. Chem. 282, (peer-reviewed)
Pages: 27847-27856
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Stomach-specific calpain, nCL-2, localizes in mucus cells and proteolyzes the β-subunit of coatomer complex, β-COP.2006
Author(s)
Hata, S., Koyama, S., Kawahara, H., Doi, N., Maeda, T., Toyama-Sorimachi, N., Abe, K., Suzuki, K., and Sorimachi, H.
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Journal Title
J. Biol. Chem. 281, (peer-reviewed)
Pages: 11214-11224
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Proteolytic regulation of creatine-kinase by muscle-specific RING-finger protein, MURF1.2008
Author(s)
Koyama, S., Hata, S., Witt, C. C., Ono, Y., Ojima, K., Chiba, T., Doi, N., Kitamura, F., Tanaka, K., Abe, K., Witt, S., Labeit, S., and Sorimachi, H.
Organizer
Nogei-kagakukai
Place of Presentation
Nagoya, Japan
Year and Date
2008-03-28
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Possible regulation of energy metabolism by MURF1.2007
Author(s)
Koyama, S., Hata, S., Witt, C. C., Ono, Y., Ojima, K., Chiba, T., Doi, N., Kitamura, F., Tanaka, K., Abe, K., Witt, S., Labeit, S., and Sorimach, H.
Organizer
Nogei-kagakukai
Place of Presentation
Tokyo, Japan
Year and Date
2007-03-25
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Physiological functions of a stomach-specific calpain, nCL-2, in the mucus secreting cells.2006
Author(s)
ata, S., Koyama, S., Kawahara, H., Doi, N., Maeda, T., Toyama-Sorimachi, N., Abe, K., Suzuki, K., and Sorimachi, H.
Organizer
20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB Congress.
Place of Presentation
Kyoto, Japan
Year and Date
2006-06-23
Description
「研究成果報告書概要(欧文)」より
