| Project/Area Number |
12671817
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Functional basic dentistry
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| Research Institution | The Nippon Dental University |
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Principal Investigator |
SAITOH Eiichi The Nippon Dental University School of Dentistry at Niigata Department of Biochemistry, Associate Professor, 新潟歯学部, 助教授 (40120662)
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| Project Period (FY) |
2000 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2002: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2001: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2000: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | Saliva / Human cystatin / Porphyromonas gingivalis / Cvsteine proteinases / Recombinant cystatins / Cystatin superfamily / Reverse zymography / Defense molecules / 唾液 / ヒトシスタチン / リバースザイモグラフイー / 生体防御 / シスタチンC / 組換え型シスタチンS / ウナギ体表粘液 / システインプロテアーゼインヒビター / アミノ酸配列 / カルボキシメチルパパイン / プロテオーム解析 / ファミリー2シスタチン / リコンビナントシスタチン / サイトカイン誘導能 / 歯肉線維芽細胞 / モノクローナル抗体 / プロテアーゼインヒビター / 唾液タンパク質 / タンパク質工学 |
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| Research Abstract |
This research project focused on the viewpoint of practical use of biological activities of human salivary cystatins and natural cysteine proteinase inhibitors from plants and animals. In addition, a system enabling large-scale production of engineered cystatins at a Ievel of several hundreds to several thousands milligrams was investigated. Based on these concepts and plans, novel cysteine proteinase inhibitors were screened as well by the reverse zymography developed in this study. The results obtained are summarized as follows: (1) Recombinant cystatins produced by E. coli were demonstrated to induce interleukin-6 and interleukin-8 productions in the human gingival fibroblast cells via cell surface molecules. (2) The engineered salivary type cystatins were also found to depress the formation of axillary odors by microorganisms such as Brevibacterium. (3) Some experimental data have indicated that there are some components mat inhibit the growth of Porphyromonas gingivalis in the hea
… More
t-extract of the rice bran of Niigata-Koshihikari. At present, we have been studying not only oryzacystatins with molecular weight of 13 kDa but also several candidate proteins of 50-70 kDa that tightly bind to a cysteine proteinase, papain and ficin. The detail features of the substances are now under investigation by the reverse zymographical technique since rice cysteine proteinase inhibitors could be a potent inhibitor of the growth of P. gingivalis as well as salivary cystatins. (6) The mechanism of a defense system present in the skin mucus of fishes may be helpful to study physiological roles of salivary proteins in oral cavity. Hence, we have attempted to search for flie defense proteins such as proteainses, proteinase inhibitors, and the lectins in the mucus fluid of the Japanese eel. As the result, the skin mucus was shown to contain novel cysteine proteinase inhibitor, with molecular masses of about 13-15 kDa. However, it is obscure at present that the inhibitors may be the members of cystatin superfamily, or not. Taken together, it can be concluded that cysteine proteinase inhibitors from plants and animals are thought to be a considerably useful biomaterial for the maintenance of oral health and for the clinical trials of periodontitis. Less
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