绿豆（Phaseolus aureus）核酸酶。使用二核苷硫代磷酸酯对DNA切割反应的机理研究。
Mung bean (Phaseolus aureus) nuclease. A mechanistic investigation of the DNA-cleavage reaction using a dinucleoside phosphorothioate.
作者信息
Hamblin M R, Cummins J H, Potter B V
出版信息
Biochem J. 1987 Feb 1;241(3):827-33. doi: 10.1042/bj2410827.
Abstract
Mung-bean (Phaseolus aureus) nuclease has been found to cleave the Sp diastereoisomer of 5'-O-thymidyl 3'-O-(2'-deoxyadenosyl)phosphorothioate, (Sp)-d[Ap(S)T], in 18O-labelled water with inversion of configuration at phosphorus to give (Sp)-thymidine 5'-[16O, 18O]phosphorothioate, the stereochemistry of which was deduced by methylation to (Rp,Sp)-thymidine 5'-S-methyl-O-methyl-[16O,18O]phosphorothioate and 31P-n.m.r. analysis. This result is consistent with a mechanism involving a direct 'in-line' attack of water on DNA for the nuclease-catalysed reaction without the involvement of a covalent nucleotidylated-enzyme intermediate.
摘要
已发现绿豆核酸酶能在18O标记的水中切割5'-O-胸苷基3'-O-(2'-脱氧腺苷基)硫代磷酸酯的Sp非对映异构体(Sp)-d[Ap(S)T]，磷原子处构型翻转，生成(Sp)-胸苷5'-[16O, 18O]硫代磷酸酯，其立体化学结构通过甲基化转化为(Rp,Sp)-胸苷5'-S-甲基-O-甲基-[16O,18O]硫代磷酸酯并经31P核磁共振分析推导得出。该结果与一种机制相符，即对于核酸酶催化的反应，水直接“直线式”进攻DNA，不涉及共价核苷酸化酶中间体。
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