1999 Fiscal Year Final Research Report Summary
Studies on Production of Human Salivary Type Cystatins by Genetic Engineering and their Application to Dental Medicine
| Project/Area Number |
09671910
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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 | Nippon Dental University |
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Principal Investigator |
SAITOH Eiichi Nippon Dental University, Department of Oral Biochemistry, Associate Professor, 新潟歯学部, 助教授 (40120662)
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| Project Period (FY) |
1997 – 1999
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| Keywords | Saliva / Cysteine proteinase inhibitor / Recombinat cystatin / Chimeric cystatin / Site directed mutagenesis / Utilization of recombinant cystatin / Protein engineering / Structural and functional relationship of protein |
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| Research Abstract |
The head investigator took the focus to human cystatins (cystatin S, cystatin SA, systatin SN, cystatin C, and cystatin D), noncompetitive inhibitors for cysteine proteinases of the papain family, in order to utilize their physiological functions. In the first stage of this research project, the E. coli system for the large-scale production of cystatins S. SN, and SA was developed. The engineered cystatins created by this system were considered to be equivalent to the natural cystatins. This system was also employed for the production of cystatin variants and chimeric cystatins between cystatin S and cystatin C. The kinetic study with above cystatins and minicking peptides showed that the first and second hairpin loops of the cystatins are important in the inhibition of cysteine proteinases. The success in creation of chimeric cystatins suggested that it is possible to design and produce artificial cystatins by protein engineering.
The attempts for seeking ways to utilize recombinant human cystatins were finally made. As the results, the cystatins were found to inhibit the growth of Porphyromonas gingivalis and to induce interleukins (IL-6 and IL-8) productions by human gingival fibroblasts. In additions, the cystatins were clearly demonstrated to be a strong inhibitor for cathepsin K, which is associated with osteoclastic born resorpiton. These findings emphasize that the cystatins possess possibility of use in therapy for osteoporosis and periodontal disease.
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