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Investigation of physiological functions of a 3-mercaptopyruvate sulfurtransferase

3-巯基丙酮酸硫转移酶的生理功能研究

基本信息

批准号：
07672394
负责人：
ISHII Kazuyuki
金额：
$ 1.41万
依托单位：
MEIJI COLLEGE OF PHARMACY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07672394/
关键词：
human genome cloning cDNA cloning 3-mercaptopyruvate sulfurtransferase rhodanese expression

项目摘要

1. A complete amino acid structure of human liver 3-mercaptopyruvate sulfurtransferase (3-MST,EC2.8.1.2) was determined by sequence analysis of cDNA and purified enzyme. The enzyme consists of 297-amino acid residues with a calculated molecular mass of 33,176.7daltons. Sequence identity in cDNA and the deduced amino acid sequence are 66.9 and 59,7% respectivity, between human 3-MST and rhodanese. By their entire sequence similarity 3-MST and rhodanese are confirmed to be evolutionarily related enzyme. When the 3-MST cDNA was transiently expressed in Escherchia coli and Cos7 cells, the 3-MST activity increased 20-fold and 45-fold, respectively.2. cDNA for the human rhodanese (thiosulfate ; cyanide sulfurtransuferase, EC 2.8.1.1) was cloned from a human fetal liver cDNA library. Sequencing of the cDNA revealed an open reading frame that encodes a 297-residue polypeptide with a calculated mass of 33427 daltons. When the rhodanese cDNA was transiently expressed in Escherchia coli and Cos7 cells, the rhodanese activity increased 40-fold and 150-fold, respectively. Sequence homology analysis showed that the human rhodanese is 89.6% identical to bovine, 90.2% identical to rat, 91.2% identical to mouse and Chinese hamster, and 71.4% similar to avian counterparts, respectively, and that rhodanese was highly conserved across evolution.

1.通过对人肝3-巯基丙酮酸硫转移酶（3-MST，EC2.8.1.2）cDNA和纯化酶的序列分析，确定了其完整的氨基酸结构。该酶由297个氨基酸残基组成，计算分子量为33，176.7道尔顿。人3-MST与Rhodanese的cDNA序列和推导的氨基酸序列的同源性分别为66.9%和59.7%。3-MST和Rhodanese的全序列相似性证实它们是进化上相关的酶。将3-MST cDNA在大肠杆菌和Cos 7细胞中瞬时表达后，3-MST活性分别提高了20倍和45倍.从人胎肝cDNA文库中克隆人罗丹酸（硫代硫酸盐;氰化物硫转移酶，EC 2.8.1.1）的cDNA。cDNA序列分析表明，该基因的开放阅读框编码297个氨基酸残基的多肽，计算质量为33427道尔顿。当rhodanese的cDNA在土方大肠杆菌和Cos 7细胞中瞬时表达时，rhodanese活性分别增加了40倍和150倍。序列同源性分析表明，人的Rhodanese与牛的同源性为89.6%，与大鼠的同源性为90.2%，与小鼠和中国仓鼠的同源性为91.2%，与鸟类的同源性为71.4%，并且Rhodanese在进化过程中高度保守。