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Biochem J. 1996 Feb 15;314 ( Pt 1)(Pt 1):321-6. doi: 10.1042/bj3140321.
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S-adenosyl-L-methionine-dependent methyl transfer: observable precatalytic intermediates during DNA cytosine methylation.S-腺苷-L-甲硫氨酸依赖性甲基转移：DNA胞嘧啶甲基化过程中可观察到的催化前中间体。
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Structure of the Q237W mutant of HhaI DNA methyltransferase: an insight into protein-protein interactions.HhaI DNA甲基转移酶Q237W突变体的结构：对蛋白质-蛋白质相互作用的深入了解。
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The small subunit of M. AquI is responsible for sequence-specific DNA recognition and binding in the absence of the catalytic domain.嗜盐栖热栖热放线菌M. AquI的小亚基在缺乏催化结构域的情况下负责序列特异性DNA识别和结合。
J Bacteriol. 2003 Feb;185(4):1284-8. doi: 10.1128/JB.185.4.1284-1288.2003.

本文引用的文献

1
Crystal structure of the HhaI DNA methyltransferase complexed with S-adenosyl-L-methionine.与S-腺苷-L-甲硫氨酸复合的HhaI DNA甲基转移酶的晶体结构。
Cell. 1993 Jul 30;74(2):299-307. doi: 10.1016/0092-8674(93)90421-l.
2
HhaI methyltransferase flips its target base out of the DNA helix.HhaI甲基转移酶将其靶碱基翻转出DNA螺旋。
Cell. 1994 Jan 28;76(2):357-69. doi: 10.1016/0092-8674(94)90342-5.
3
The DNA (cytosine-5) methyltransferases.DNA（胞嘧啶-5）甲基转移酶
Nucleic Acids Res. 1994 Jan 11;22(1):1-10. doi: 10.1093/nar/22.1.1.
4
Three-dimensional structure of the adenine-specific DNA methyltransferase M.Taq I in complex with the cofactor S-adenosylmethionine.腺嘌呤特异性DNA甲基转移酶M.Taq I与辅因子S-腺苷甲硫氨酸结合的三维结构。
Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):10957-61. doi: 10.1073/pnas.91.23.10957.
5
Functional analysis of Gln-237 mutants of HhaI methyltransferase.HhaI甲基转移酶Gln-237突变体的功能分析
Nucleic Acids Res. 1995 Feb 25;23(4):620-7. doi: 10.1093/nar/23.4.620.
6
A bacterial methyltransferase M.EcoHK311 requires two proteins for in vitro methylation.一种细菌甲基转移酶M.EcoHK311在体外甲基化过程中需要两种蛋白质。
Nucleic Acids Res. 1995 Jan 11;23(1):103-8. doi: 10.1093/nar/23.1.103.
7
Selfish behavior of restriction-modification systems.限制修饰系统的自私行为。
Science. 1995 Feb 10;267(5199):897-9. doi: 10.1126/science.7846533.
8
The crystal structure of HaeIII methyltransferase convalently complexed to DNA: an extrahelical cytosine and rearranged base pairing.与DNA共价结合的HaeIII甲基转移酶的晶体结构：一个螺旋外胞嘧啶和重排的碱基配对。
Cell. 1995 Jul 14;82(1):143-53. doi: 10.1016/0092-8674(95)90060-8.
9
Kinetic and catalytic mechanism of HhaI methyltransferase.HhaI甲基转移酶的动力学和催化机制。
J Biol Chem. 1987 Apr 5;262(10):4778-86.
10
Cloning and sequencing of a cDNA encoding DNA methyltransferase of mouse cells. The carboxyl-terminal domain of the mammalian enzymes is related to bacterial restriction methyltransferases.小鼠细胞DNA甲基转移酶编码cDNA的克隆与测序。哺乳动物酶的羧基末端结构域与细菌限制性甲基转移酶相关。
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具有两条多肽链的细菌甲基转移酶M.EcoHK31I的过量表达、纯化及特性分析

Overproduction, purification and characterization of M.EcoHK31I, a bacterial methyltransferase with two polypeptides.

作者信息

Lee K F, Liaw Y C, Shaw P C

机构信息

Department of Biochemistry, The Chinese University of Hong Kong.

出版信息

Biochem J. 1996 Feb 15;314 ( Pt 1)(Pt 1):321-6. doi: 10.1042/bj3140321.

DOI:10.1042/bj3140321
PMID:8660301
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1217043/
Abstract

The two overlapping genes coding for EcoHK31I methyltransferase have previously been cloned, sequenced and expressed [Lee, Kam and Shaw (1995) Nucleic Acids Res. 23, 103-108]. Here we describe protocols developed to purify polypeptides alpha and beta together or separately, to apparent homogeneity by various chromatographic media. M.EcoHK31I is a heterodimer with a native molecular mass of 61 kDa. Its specific activity towards non-methylated lambda DNA was 3.0 x 10(5) units per mg of protein. The respective denatured molecular masses of polypeptides alpha and beta were 38 and 23 kDa, and their pI values were 8.7 and 6.8. Initial rate kinetic parameters of the native enzyme were 2.0 nM, 0.58 microM and 3 min-1 for KmDNA, KmAdoMet and kcat. respectively, where AdoMet stands for S-adenosyl-L-methionine. Fully active enzyme was reconstituted by co-purifying the two separately synthesized polypeptides, and activity assays confirmed our previous finding that two polypeptides were needed to methylate substrate DNA.

摘要

编码EcoHK31I甲基转移酶的两个重叠基因先前已被克隆、测序和表达[Lee、Kam和Shaw（1995年）《核酸研究》23卷，第103 - 108页]。在此，我们描述了所开发的方案，通过各种色谱介质一起或分别纯化α和β多肽，直至达到明显的均一性。M.EcoHK31I是一种天然分子量为61 kDa的异二聚体。其对未甲基化的λDNA的比活性为每毫克蛋白质3.0×10⁵单位。α和β多肽各自的变性分子量分别为38 kDa和23 kDa，其pI值分别为8.7和6.8。天然酶的初始速率动力学参数，对于KmDNA、KmAdoMet和kcat分别为2.0 nM、0.58 μM和3 min⁻¹，其中AdoMet代表S - 腺苷 - L - 甲硫氨酸。通过共纯化两种分别合成的多肽重构了完全活性的酶，活性测定证实了我们先前的发现，即需要两种多肽来甲基化底物DNA。