磷酸脱氧核糖基转移酶，设计用于合成脱氧核苷酸的酶。
Phosphodeoxyribosyltransferases, designed enzymes for deoxyribonucleotides synthesis.
机构信息
Institut Pasteur, Unité de Chimie et Biocatalyse, CNRS, UMR 3523, 75724 Paris cedex 15, France.
出版信息
J Biol Chem. 2013 Mar 1;288(9):6534-41. doi: 10.1074/jbc.M112.446492. Epub 2013 Jan 16.
Abstract
A large number of nucleoside analogues and 2'-deoxynucleoside triphosphates (dNTP) have been synthesized to interfere with DNA metabolism. However, in vivo the concentration and phosphorylation of these analogues are key limiting factors. In this context, we designed enzymes to switch nucleobases attached to a deoxyribose monophosphate. Active chimeras were made from two distantly related enzymes: a nucleoside deoxyribosyltransferase from lactobacilli and a 5'-monophosphate-2'-deoxyribonucleoside hydrolase from rat. Then their unprecedented activity was further extended to deoxyribose triphosphate, and in vitro biosyntheses could be successfully performed with several base analogues. These new enzymes provide new tools to synthesize dNTP analogues and to deliver them into cells.
摘要
已经合成了大量的核苷类似物和 2'-脱氧核苷三磷酸（dNTP）来干扰 DNA 代谢。然而，在体内，这些类似物的浓度和磷酸化是关键的限制因素。在这种情况下，我们设计了酶来切换连接到脱氧核糖一磷酸上的碱基。活性嵌合体是由两种亲缘关系较远的酶制成的：一种来自乳杆菌的核苷脱氧核糖基转移酶和一种来自大鼠的 5'-单磷酸-2'-脱氧核糖核苷水解酶。然后，它们以前所未有的活性进一步扩展到脱氧核糖三磷酸，并成功地在体外生物合成了几种碱基类似物。这些新的酶为合成 dNTP 类似物并将其递送到细胞内提供了新的工具。
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