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

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cDNA和纯化的酶的序列分析确定人肝3-丙型丙酮酸硫酸盐硫酸盐硫酸盐的完整氨基酸结构（3-MST，EC2.8.1.2）。该酶由297-氨基酸残基组成，计算出的分子质量为33,176.7daltons。在人类3-MST和Rhodanese之间，cDNA和推导的氨基酸序列的序列身份为66.9和59.7％。通过他们的整个序列相似性，第3-MST和罗丹尼人被证实是进化相关的酶。当在大肠杆菌和COS7细胞中瞬时表达3-M的cDNA时，第三个活性分别增加了20倍和45倍。2。从人类胎儿肝cDNA文库中克隆了人类罗丹尼人（硫代硫酸盐；氰化物硫硫酸盐，EC 2.8.1.1）的cDNA。 cDNA的测序揭示了一个开放的阅读框架，该框架编码了297个残留的多肽，其计算质量为33427 daltons。当罗达氏c cDNA在大肠杆菌和COS7细胞中瞬时表达时，罗丹斯活性分别增加了40倍和150倍。序列同源性分析表明，人类罗丹尼人与牛相同89.6％，90.2％与大鼠相同，91.2％与小鼠和中国仓鼠相同，分别为71.4％，分别与禽类相似，而罗丹尼人在进化过程中高度保存。