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首页> 外文期刊>Chemistry: A European journal >Polymerase-Catalysed Incorporation of Glucose Nucleotides into a DNA Duplex
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Polymerase-Catalysed Incorporation of Glucose Nucleotides into a DNA Duplex

机译：聚合酶催化将葡萄糖核苷酸掺入DNA双链体

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摘要

The enzymatic recognition of six-membered ring nucleoside triphosphates— in particular the 6’-triphosphates of (b-d-glucopyranosyl)thymine, (2’,3’-dideoxy-b-d-glucopyranosyl)thymine, (3’,4’-dideoxy-b-d-glucopyranosyl) thymine and (2’,3’-dideoxy-b-d-glucopyranosyl) adenine—was investigated. Despite the facts that the pyranose nucleic acids obtained by polymerisation of these monomers do not hybridise in solution with DNA and that the geometry of a DNA strand in a natural duplex differs from that of a pyranose nucleic acid, elongation of the DNA duplex with all four nucleotide analogues by Vent (exo) polymerase was observed. Modelling experiments showed that hydrogen bonds are formed when 2’,3’-dideoxy-b-homo- T building blocks or b-d-gluco-T building blocks are incorporated opposite adenosine residues in the template but not when they are incorporated opposite thymine residues in the template. The model shows a near perfect alignment of a secondary hydroxy group at the end of the primer and the a-phosphate group of the incoming triphosphate. The results of these experiments provide new information on the role of the active site of the enzyme in the polymerisation reaction.
机译：六元环核苷三磷酸的酶促识别-特别是（bd-吡喃葡萄糖基）胸腺嘧啶，（2'，3'-二脱氧-bd-吡喃葡萄糖基）胸腺嘧啶，（3'，4'-二脱氧-研究了bd-吡喃葡萄糖基）胸腺嘧啶和（2'，3'-二脱氧-bd-吡喃葡萄糖基）腺嘌呤。尽管通过聚合这些单体获得的吡喃糖核酸不会在溶液中与DNA杂交，并且天然双链体中DNA链的几何结构与吡喃糖核酸不同，但DNA双链体在所有四个碱基上的延伸观察到Vent（exo）聚合酶的核苷酸类似物。建模实验表明，当2'，3'-dideoxy-b-homo-T构件或bd-gluco-T构件在模板中与腺苷残基相反时形成氢键，而在2'，3'-dideoxy-b-homo-T构件中与腺苷残基相反时，则不形成氢键。模板。该模型显示了底漆末端的仲羟基与引入的三磷酸的α-磷酸基团几乎完全对准。这些实验的结果提供了有关酶的活性位点在聚合反应中的作用的新信息。

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