Identification and phenotypic analysis of two glyoxalase II encoding genes from Saccharomyces cerevisiae, GLO2 and GLO4, and intracellular localization of the corresponding proteins
Identification and phenotypic analysis of two glyoxalase II encoding genes from Saccharomyces cerevisiae, GLO2 and GLO4, and intracellular localization of the corresponding proteins
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DOI：
10.1074/jbc.272.34.21509
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发表时间：
1997-08-22
影响因子：
4.8
通讯作者：
Breitenbach, M
中科院分区：
文献类型：
--
作者：
Bito, A;Haider, M;Breitenbach, M
我们从酿酒酵母中分离并鉴定了两个乙醛酸酶II基因，GLO2基因编码区对应一个274个氨基酸的蛋白质，分子质量为31,306个道尔顿，GLO4基因的开放阅读框可翻译成一个285个氨基酸的蛋白质，分子质量为32,325个道尔顿，推导出的蛋白质的氨基酸序列与人乙醛酸酶II的序列有59.1%的同源性，在以葡萄糖或甘油为碳源的情况下，GLO2/GLO4双缺失菌株完全不含乙二醛酶II活性，在植物生长过程中没有明显的表型。该菌株对外源甲基乙二醛表现出与乙二醇酶I缺陷型菌株相似的高敏感性，而GLO2基因在葡萄糖和甘油上都表达，GLO4基因只在甘油上表达，活性Glo2p蛋白定位在细胞质中，活性Glo4p蛋白定位在线粒体基质中。Glo2全长编码区在大肠杆菌中的异源表达产生了一个活性蛋白，而要在大肠杆菌中获得一个活性的Glo4p蛋白，必须通过缩短GLO4开放阅读框的5‘端来去除N端的线粒体转运肽。
We have isolated and characterized two genes coding for the glyoxalase II enzyme from Saccharomyces cerevisiae, The coding region of the GLO2 gene corresponds to a protein with 274 amino acids and a molecular mass of 31,306 daltons, The open reading frame of the GLO4 gene could be translated into a protein with 285 amino acids and a molecular mass of 32,325 daltons, The amino acid sequences of the deduced proteins are 59.1% identical and show high similarities to the sequence of the human glyoxalase II, When grown on either glucose or glycerol as a carbon source, a glo2 glo4 double deletion strain contains no glyoxalase II activity at all and shows no obvious phenotype during vegetative growth, However, this strain showed a similar high sensitivity against exogenous methylglyoxal as compared with a glyoxalase I-deficient strain, Whereas the GLO2 gene is expressed on both glucose and glycerol, the GLO4 gene is only active on glycerol, The active Glo2p protein is localized in the cytoplasm and the active Glo4p in the mitochondrial matrix. Heterologous expression of the full-length GLO2 coding region in Escherichia coli resulted in an active protein, However, to get an active Glo4p protein in E. coli, the putative mitochondrial transit peptide at the N-terminal end had to be removed by shortening the 5' end of the GLO4 open reading frame.