Studies on Regulation of Cellular Function by Calpastatin

钙蛋白酶抑制剂调节细胞功能的研究

基本信息

批准号：
01580157
负责人：
MAKI Masatoshi
金额：
$ 1.28万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1991
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01580157/
关键词：
Calpain Calpastatin Proteinase Proteinase Inhibitor Protein Kinase C calcium channel プロテア-ゼ

项目摘要

Calpastatin is endogenous inhibitor protein acting specifically on intracellular calcium dependent proteinase(calpain). In the present study we extended structure-function analyses of calpastatin and examined roles of this inhibitor in animal cells. The major results obtained were as follows :1. Locus of human calpastatin gene was assigned to chromosome 5ql4-22.2. Highly conserved regions among the four internally repetitive domains were separated by introns in the genomic DNA.3. The 27-residue synthetic oligopeptide corresponding to'the Exon 1B was active as calpain inhibitor, and it suppressed TPA induced down-regulation of protein kinase C when added to culture medium of BIM cells(neuroblastoma cell line).4.2-dimensional NMR studies have revaeled that the oligopeptide forms type I beta-turn structure in the highly conserved sequence.5. Monoclonal antibodies were prepared for human calpastatin, and a method of distinguishing two types(muscle and erythrocyte)of calpastatin molecules were developed.In contrast to mammalian erythrocytes, chicken erythrocytes(nucleated cells)were suggested to retain amino-terminal domains of calpastatin and to have a muscle type structure.6. MRNA and protein levels of m-calpain and calpastatin were found to be elevated in HTLV-I infected T-cell leukemia cell lines, but not in HTLV-I tax-transformed fibroblast cell lines.7. Besides calpain inhibition, calpastatin was suggested to have L-type calcium channel activating function.

CALPASTATIN是内源性抑制剂蛋白，该抑制剂专门作用于细胞内钙依赖性蛋白酶（CALPAIN）。在本研究中，我们扩展了Calpastatin的结构 - 功能分析，并检查了该抑制剂在动物细胞中的作用。获得的主要结果如下：1。人calpastatin基因的基因座分配给5ql4-22.2染色体。基因组DNA中的内含子分离了四个内部重复域中的高度保守区域。3。 The 27-residue synthetic oligopeptide corresponding to'the Exon 1B was active as calpain inhibitor, and it suppressed TPA induced down-regulation of protein kinase C when added to culture medium of BIM cells(neuroblastoma cell line).4.2-dimensional NMR studies have revaeled that the oligopeptide forms type I beta-turn structure in the highly conserved sequence.5.为人钙蛋白酶制备了单克隆抗体，并开发了一种区分两种类型的calpastatin分子（肌肉和红细胞）的方法。与哺乳动物红细胞相反，建议养鸡红细胞（鸡肉红细胞）（成核细胞）保持乳腺氨基 - 氨基氨基氨基氨基含量的类型和calpaspastation的幼稚犬脉络脉络脉络素的6.发现在HTLV-I感染的T细胞白血病细胞系中，M-calpain和Calpastatin的mRNA和蛋白质水平升高，但在HTLV-I税收转换的成纤维细胞系中却没有升高。7。除钙蛋白酶抑制外，建议钙蛋白酶具有L型钙通道激活功能。