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Kinetic analysis of an efficient DNA-dependent TNA polymerase
其他题名Kinetic analysis of an efficient DNA-dependent TNA polymerase
Allen Horhota; Keyong Zou; Justin K. Ichida; Yu B(俞飚)
2005
发表期刊J. Am. Chem. Soc.
ISSN0002-7863
卷号127期号:20页码:7427-7434
摘要alpha-L-Threofuranosyl nucleoside triphosphates (tNTPs) are tetrafuranose nucleoside derivatives and potential progenitors of present-day beta-D-2'-deoxyribofuranosyl nucleoside triphosphates (dNTPs). Therminator DNA polymerase, a variant of the 9 degrees N DNA polymerase, is an efficient DNA-directed threosyl nucleic acid (TNA) polymerase. Here we report a detailed kinetic comparison of Therminator-catalyzed TNA and DNA syntheses. We examined the rate of sing le-nucleotide incorporation for all four tNTPs and dNTPs from a DNA primer-template complex and carried out parallel experiments with a chimeric DNA-TNA primer-DNA template containing five TNA residues at the primer X-terminus. Remarkably, no drop in the rate of TNA incorporation was observed in comparing the DNA-TNA primer to the all-DNA primer, suggesting that few primer-enzyme contacts are lost with a TNA primer. Moreover, comparison of the catalytic efficiency of TNA synthesis relative to DNA synthesis at the downstream positions reveals a difference of no greater than 5-fold in favor of the natural DNA substrate. This disparity becomes negligible when the TNA synthesis reaction mixture is supplemented with 1.25 mM MnCl2. These results indicate that Therminator DNA polymerase can recognize both a TNA primer and tNTP substrates and is an effective catalyst of TNA polymerization despite changes in the geometry of the reactants.
学科领域生命有机化学
部门归属Boston College ; Howard Hughes Medical Institute and Massachusetts General Hospital; 中科院上海有机所
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收录类别SCI
语种英语
WOS记录号WOS:000229244600047
引用统计
文献类型期刊论文
条目标识符http://ir.sioc.ac.cn/handle/331003/19009
专题生命有机化学国家重点实验室
通讯作者Yu B(俞飚)
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GB/T 7714
Allen Horhota,Keyong Zou,Justin K. Ichida,et al. Kinetic analysis of an efficient DNA-dependent TNA polymerase[J]. J. Am. Chem. Soc.,2005,127(20):7427-7434.
APA Allen Horhota,Keyong Zou,Justin K. Ichida,&俞飚.(2005).Kinetic analysis of an efficient DNA-dependent TNA polymerase.J. Am. Chem. Soc.,127(20),7427-7434.
MLA Allen Horhota,et al."Kinetic analysis of an efficient DNA-dependent TNA polymerase".J. Am. Chem. Soc. 127.20(2005):7427-7434.
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