Kinetic analysis of an efficient DNA-dependent TNA polymerase

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	Kinetic analysis of an efficient DNA-dependent TNA polymerase
	Allen Horhota ; Keyong Zou ; Justin K. Ichida ; Yu B(俞飚)
刊名	J. Am. Chem. Soc.
	2005
卷号	127 期号:20 页码:7427-7434
ISSN号	0002-7863
其他题名	Kinetic analysis of an efficient DNA-dependent TNA polymerase
通讯作者	俞飚
英文摘要	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.
学科主题	生命有机化学
收录类别	SCI
原文出处	http://dx.doi.org/10.1021/ja0428255
语种	英语
WOS记录号	WOS:000229244600047
公开日期	2013-02-25
内容类型	期刊论文
源URL	[http://202.127.28.38/handle/331003/19009]
专题	上海有机化学研究所_生命有机化学国家重点实验室
推荐引用方式 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.