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Investigation of the unique character of the squid novel neutral metalloprotease

鱿鱼新型中性金属蛋白酶的独特性质研究

基本信息

批准号：
12660191
负责人：
TSUCHIYA Takahide
金额：
$ 2.62万
依托单位：
Sophia 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 2001
项目状态：
已结题

项目摘要

The metallprotease which has activity in the neutral found out by only the squid muscle exists. This neutral metallprotease shows high activity to myosin, and was considered to be the enzyme related to the metabolism of muscular protein. However, although activity was accepted in TEUTHOIDEA squid muscles, such as a squid, was not activity in the muscles of SEPIOIDEA. Furthermore, what has functions other than line protein metabolism in it since activity is high to the very few liver of the amount of myosin was conjectured. Three kinds of isoform from which the decomposition position of myosin differs existed in this enzyme. To clear the function of this novel protease, the determination of all base arrangement was tried by the PCR method. When amino acid arrangement was presumed base arrangement, it turns out by the squid and the spear squid that the methods of the existence of isoform differ. The precursor structure where any isoform had a pro-domain was taken, and it became clear that it is protease of secretion nature with signal peptide. It was able to be shown clearly from base arrangement that it belongs to an astacin family again. The thing in connection with as digestive enzyme regulation and generating of a growth factor in the protease belonging to an astacin family etc. exists. When the phylogenetic tree of a protease domain was investigated, it also turns out that the protease of a squid does not belong to the cluster of which astacin family known conventionally. Although the novel protease of this squid was before named myosinase, it changed with astacin-like squid metalloprotease.

在中性条件下具有活性的金属蛋白酶仅在鱿鱼肌肉中发现。这种中性金属蛋白酶对肌球蛋白有很高的活性，被认为是与肌肉蛋白质代谢有关的酶。然而，虽然活动被接受在TEUTHOIDEA鱿鱼肌肉，如鱿鱼，是不是活动在SEPIOIDEA的肌肉。此外，由于肌球蛋白的量的极少数肝脏活性高，因此其具有除线蛋白代谢以外的功能。该酶存在三种亚型，其肌球蛋白的分解位置不同。为了明确该蛋白酶的功能，我们尝试用PCR方法对该蛋白酶的全碱基排列进行检测。当氨基酸排列被假定为碱基排列时，鱿鱼和矛鱿鱼发现同种型存在的方法不同。获得了其中任何同种型具有前结构域的前体结构，并且变得清楚的是，它是具有信号肽的分泌性质的蛋白酶。从碱基排列上可以清楚地表明它又属于一个astacin家族。存在与作为消化酶的调节和属于虾青素家族等的蛋白酶中的生长因子的产生有关的物质。当研究蛋白酶结构域的系统发育树时，还发现鱿鱼的蛋白酶不属于常规已知的astacin家族的簇。虽然这种鱿鱼的新蛋白酶以前被命名为肌球蛋白酶，但它变成了虾红素样鱿鱼金属蛋白酶。