拟南芥硫酯酶乙二醛酶II的分子克隆与特性分析
Molecular cloning and characterization of the thiolesterase glyoxalase II from Arabidopsis thaliana.
作者信息
Ridderström M, Mannervik B
机构信息
Department of Biochemistry, Uppsala University, Biomedical Center, Sweden.
出版信息
Biochem J. 1997 Mar 1;322 ( Pt 2)(Pt 2):449-54. doi: 10.1042/bj3220449.
cDNA encoding glyoxalase II from Arabidopsis thaliana has been cloned and sequenced. The isolated 894 bp segment included a sequence of 774 bp encoding a protein with a calculated molecular mass of 28,791 Da. The amino acid sequence deduced from the A. thaliana cDNA showed 54% identity with that of the human enzyme. Searches in databanks identified seven additional DNA sequences from different species with high similarity to glyoxalase II. Certain limited regions, one rich in histidine residues, shared 100% identity. A 29 kDa protein with an isoelectric point of 6.2 was obtained by heterologous expression of the A. thaliana cDNA in Escherichia coli. Homogeneous enzyme was obtained by affinity purification and its catalytic parameters with thiolesters of glutathione were similar to those for human glyoxalase II. The structural and functional similarities between glyoxalase II from A. thaliana and from human tissues suggest a common evolutionary origin.
已克隆并测序了拟南芥中编码乙二醛酶II的cDNA。分离出的894 bp片段包含一个774 bp的序列，该序列编码一种计算分子量为28,791 Da的蛋白质。从拟南芥cDNA推导的氨基酸序列与人类酶的氨基酸序列具有54%的同一性。数据库搜索鉴定出另外七个来自不同物种的与乙二醛酶II高度相似的DNA序列。某些有限区域，其中一个富含组氨酸残基，具有100%的同一性。通过在大肠杆菌中异源表达拟南芥cDNA获得了一种等电点为6.2的29 kDa蛋白质。通过亲和纯化获得了均一的酶，其与谷胱甘肽硫酯的催化参数与人类乙二醛酶II的催化参数相似。拟南芥和人类组织中的乙二醛酶II之间的结构和功能相似性表明它们有共同的进化起源。
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