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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

本研究项目主要关注人唾液半胱氨酸蛋白酶抑制剂和动植物天然半胱氨酸蛋白酶抑制剂生物活性的实际应用。此外，研究了能够大规模生产数百至数千毫克水平的工程化半胱氨酸蛋白酶抑制剂的系统。基于这些概念和计划，新的半胱氨酸蛋白酶抑制剂筛选以及反向酶谱法在这项研究中发展。获得的结果如下：（1）大肠杆菌表达的重组半胱氨酸蛋白酶抑制剂;大肠杆菌通过细胞表面分子诱导人牙龈成纤维细胞产生白细胞介素-6和白细胞介素-8。(2)还发现工程化的唾液型半胱氨酸蛋白酶抑制剂抑制微生物如短杆菌属（Brevibacterium）的腋窝气味的形成。(3)一些实验数据表明，在牙龈卟啉单胞菌（Porphyromonasgingivalis）的生长抑制剂中， ...更多信息新潟越光米糠提取物。目前，我们不仅研究了分子量为13 kDa的水稻晶体蛋白，还研究了几种分子量为50-70 kDa的候选蛋白，它们与半胱氨酸蛋白酶、木瓜蛋白酶和无花果蛋白酶紧密结合。由于水稻半胱氨酸蛋白酶抑制剂可能是牙龈卟啉单胞菌和唾液半胱氨酸蛋白酶抑制剂生长的有效抑制剂，因此目前正在通过反向酶谱技术研究这些物质的详细特征。(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生理作用。因此，我们试图在日本鳗鲡粘液中寻找诸如蛋白酶、蛋白酶抑制剂和凝集素等蝇类防御蛋白。结果显示，皮肤粘液中含有新型半胱氨酸蛋白酶抑制剂，分子量约为13-15 kDa。然而，目前尚不清楚这些抑制剂是否是胱抑素超家族成员。两者合计，可以得出结论，半胱氨酸蛋白酶抑制剂从植物和动物被认为是一个相当有用的生物材料，用于维护口腔健康和牙周炎的临床试验。少