Studies on the production and practical use of engineered human salivary type cystatins.

工程人唾液型半胱氨酸蛋白酶抑制剂的生产和实际应用研究。

基本信息

批准号：
12671817
负责人：
SAITOH Eiichi
金额：
$ 2.11万
依托单位：
The Nippon Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

项目摘要

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

该研究项目的重点是对人类唾液性结尾蛋白的生物活性和来自植物和动物的天然半胱氨酸蛋白酶抑制剂的生物学活性的观点。此外，研究了一个系统，可在数百至数千毫克的IEVEL上大规模生产工程性半胱氨酸蛋白酶。基于这些概念和计划，这项研究中开发的反向酶学也筛选了新型半胱氨酸蛋白酶抑制剂。所获得的结果总结如下：（1）大肠杆菌产生的重组半胱氨酸蛋白酶被证明可以通过细胞表面分子诱导人牙龈成纤维细胞中的白介素-6和白介素-8。（2）还发现工程化的唾液型胱抑素可通过微生物（例如Brevibacterium）降低腋窝气味。（3）一些实验数据表明，有些成分垫抑制了HEA中卟啉念珠菌的生长……更多的niigata-koshihikari大米麸皮提取。目前，我们不仅研究了13 kDa的分子量的Oryzacystatin，而且还研究了50-70 kDa的几种候选蛋白，它们与半胱氨酸蛋白酶，木瓜蛋白酶和富菌素紧密结合。现在，该物质的细节特征正在通过反向酶学技术进行研究，因为水稻半胱氨酸蛋白酶抑制剂可能是牙龈疟原虫生长以及唾液性半胱氨酸蛋白酶生长的潜在抑制剂。（6）鱼类皮肤粘液中存在的防御系统的机制可能有助于研究唾液蛋白在口腔中的身体作用。因此，我们试图寻找蛋白酶，例如蛋白酶，蛋白酶抑制剂以及日本鳗鱼粘液液中的讲座。结果，皮肤粘液被证明包含新型半胱氨酸蛋白酶抑制剂，分子量约为13-15 kDa。但是，目前晦涩难懂的是，抑制剂可能是半胱氨酸蛋白酶超家族的成员。综上所述，可以得出结论，植物和动物的半胱氨酸蛋白酶抑制剂被认为是维持口腔健康和牙周炎临床试验的方便生物材料。较少的