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DOI	10.1073/pnas.1811518115
	Snapshots of a modified nucleotide moving through the confines of a DNA polymerase
	Kropp H.M.; Dürr S.L.; Peter C.; Diederichs K.; Marx A.
发表日期	2018
ISSN	0027-8424
起始页码	9992
结束页码	9997
卷号	115 期号:40
英文摘要	DNA polymerases have evolved to process the four canonical nucleotides accurately. Nevertheless, these enzymes are also known to process modified nucleotides, which is the key to numerous core biotechnology applications. Processing of modified nucleotides includes incorporation of the modified nucleotide and postincorporation elongation to proceed with the synthesis of the nascent DNA strand. The structural basis for postincorporation elongation is currently unknown. We addressed this issue and successfully crystallized KlenTaq DNA polymerase in six closed ternary complexes containing the enzyme, the modified DNA substrate, and the incoming nucleotide. Each structure shows a high-resolution snapshot of the elongation of a modified primer, where the modification "moves" from the 3′-primer terminus upstream to the sixth nucleotide in the primer strand. Combining these data with quantum mechanics/molecular mechanics calculations and biochemical studies elucidates how the enzyme and the modified substrate mutually modulate their conformations without compromising the enzyme's activity significantly. The study highlights the plasticity of the system as origin of the broad substrate properties of DNA polymerases and facilitates the design of improved systems. © 2018 National Academy of Sciences. All rights reserved.
英文关键词	Alkyne; Click chemistryv; Crystallography; DNA polymerase; Modified nucleotide
语种	英语
scopus关键词	DNA polymerase; nucleotide; bacterial protein; DNA; DNA directed DNA polymerase beta; Taq polymerase; 3' untranslated region; Article; biochemical analysis; chemical modification; crystallization; DNA synthesis; enzyme activity; enzyme conformation; enzyme substrate; molecular mechanics; plasticity; priority journal; quantum mechanics; structure analysis; chemistry; enzymology; molecular model; Thermus; X ray crystallography; Bacterial Proteins; Crystallography, X-Ray; DNA; DNA Polymerase I; Models, Molecular; Taq Polymerase; Thermus
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/160460
作者单位	Kropp, H.M., Department of Chemistry, Universität Konstanz, Konstanz, 78464, Germany, Konstanz Research School Chemical Biology, Universität Konstanz, Konstanz, 78464, Germany; Dürr, S.L., Department of Chemistry, Universität Konstanz, Konstanz, 78464, Germany; Peter, C., Department of Chemistry, Universität Konstanz, Konstanz, 78464, Germany, Konstanz Research School Chemical Biology, Universität Konstanz, Konstanz, 78464, Germany; Diederichs, K., Konstanz Research School Chemical Biology, Universität Konstanz, Konstanz, 78464, Germany, Department of Biology, Universität Konstanz, Konstanz, 78464, Germany; Marx, A., Department of Chemistry, Universität Konstanz, Konstanz, 78464, Germany, Konstanz Research School Chemical Biology, Universität Konstanz, Konstanz, 78464, Germany
推荐引用方式 GB/T 7714	Kropp H.M.,Dürr S.L.,Peter C.,et al. Snapshots of a modified nucleotide moving through the confines of a DNA polymerase[J],2018,115(40).
APA	Kropp H.M.,Dürr S.L.,Peter C.,Diederichs K.,&Marx A..(2018).Snapshots of a modified nucleotide moving through the confines of a DNA polymerase.Proceedings of the National Academy of Sciences of the United States of America,115(40).
MLA	Kropp H.M.,et al."Snapshots of a modified nucleotide moving through the confines of a DNA polymerase".Proceedings of the National Academy of Sciences of the United States of America 115.40(2018).