Molecular characterization of human eye and heart fatty acid ethyl ester synthase/carboxylesterase by site-directed mutagenesis.
Molecular characterization of human eye and heart fatty acid ethyl ester synthase/carboxylesterase by site-directed mutagenesis.
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通过定点诱变对人眼和心脏脂肪酸乙酯合酶/羧酸酯酶进行分子表征。
DOI：
10.1016/j.bbrc.2003.11.046
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发表时间：
2003
影响因子：
3.1
通讯作者：
Bora,NaliniS
中科院分区：
文献类型：
--
作者：
Bora,PuranS;Guruge,BandulaL;Bora,NaliniS
FAEES/羧酸酯酶以非氧化方式将酒精代谢为 FAEE，这是一种与酒精引起的终末器官损伤有关的潜在有毒分子。我们的实验室从人眼和心脏 (G12) 中分离并鉴定了 FAEES/羧酸酯酶 cDNA。为了进一步表征 G12 酶，该实验室分析了 FAEES 活性，并对 G12 的拟议活性位点进行了诱变研究。我们对 G12 酶中的 Ser 204 和 His 451 进行了突变，试图表征活性位点。我们发现这两种突变都会导致羧酸酯酶活性几乎完全丧失，而天然酶仍保持羧酸酯酶活性（G12 为 560nmol/mg 蛋白质/h）。以油酸为底物，G12的FAEES活性为170nmol/mg蛋白质/h，M204的FAEES活性为10.8nmol/mg蛋白质/h，M451的FAEES活性为8.5nmol/mg蛋白质/h。当使用乙醇作为底物时，G12的FAEES活性为240nmol/mg蛋白质/h，M204的FAEES活性为15.1nmol/mg蛋白质/h，M451的FAEES活性为6.2nmol/mg蛋白质/h。由于 G12 酶的 Ser 204 和 His 451 突变会显着降低羧酸酯酶和 FAEES 酶的活性，因此本研究表明这些残基可能对该酶的关键酶活性很重要。
The FAEES/carboxylesterase enzyme nonoxidatively metabolizes alcohol into FAEEs, potentially toxic molecules linked with alcohol-induced end-organ damage. Our laboratory has isolated and characterized a FAEES/carboxylesterase cDNA from human eye and heart (G12). In an effort to further characterize the G12 enzyme, this laboratory has analyzed the FAEES activity, as well as performed mutagenic studies on a proposed active site of G12. We have mutated Ser 204 and His 451 in the G12 enzyme in an attempt to characterize the active site. We found that both mutations cause almost total loss of carboxylesterase enzyme activity with the native enzyme remaining active for carboxylesterase activity (560nmol/mgprotein/h for G12). With oleic acid as a substrate, the FAEES activity was 170nmol/mgprotein/h for G12, 10.8nmol/mgprotein/h for M204, and 8.5nmol/mgprotein/h for M451. When ethanol was used as substrate, the FAEES activity was 240nmol/mgprotein/h for G12, 15.1nmol/mgprotein/h for M204, and 6.2nmol/mgprotein/h for M451. Since both carboxylesterase and FAEES enzyme activities are significantly lowered by mutating Ser 204 and His 451 of the G12 enzyme, this study indicates that these residues may be important for the key enzymatic activities of the enzyme.
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鉴定来自人心肌的卫星脂肪酸乙酯合酶为谷胱甘肽 S-转移酶。
DOI：
10.1172/jci114382
发表时间：
1989
期刊：
The Journal of clinical investigation
影响因子：
--
作者：
Bora,PS;Spilburg,CA;Lange,LG
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影响因子：
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影响因子：
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作者：
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影响因子：
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作者：
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影响因子：
10.8
作者：
L. D. Segel;S. Rendig;D. Mason
通讯作者：
D. Mason