Increasing DNA substrate specificity of the EcoDam DNA-(adenine N-6)-methyltransferase by site-directed mutagenesis

Elsawy H.; Chahar S.

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Increasing DNA substrate specificity of the EcoDam DNA-(adenine N-6)-methyltransferase by site-directed mutagenesis

机译：通过定点诱变提高EcoDam DNA-（腺嘌呤N-6）-甲基转移酶的DNA底物特异性

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DNA methylation catalyzed by DNA methyltransferases (MTases) is widespread in prokaryotes. In an attempt to find EcoDam variants with enhanced preference for hemimethylated DNA, the L122, P134, and V133 residues were replaced with other amino acids using site directed mutagenesis, and the catalytic activity of all variants on unmethylated and hemimethylated substrates was studied. Our results showed that, in addition to L122A, the L122S and L122A/V133L EcoDam variants were able to sense the methylation status of the 5'-GATC-3' double-stranded target recognition site and methylated only hemimethylated DNA.

机译：DNA甲基转移酶（MTase）催化的DNA甲基化在原核生物中很普遍。为了找到对半甲基化DNA的偏爱增强的EcoDam变体，使用定点诱变将L122，P134和V133残基替换为其他氨基酸，并研究了所有变体对未甲基化和半甲基化底物的催化活性。我们的结果表明，除了L122A，L122S和L122A / V133L EcoDam变体还能够感知5'-GATC-3'双链目标识别位点的甲基化状态，并且仅甲基化半甲基化的DNA。

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《Biochemistry》 |2014年第11期|共5页
作者
Elsawy H.; Chahar S.;
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正文语种 eng
中图分类生物化学;
关键词
EcoDam; rational design; protein engineering;

机译：EcoDam;合理设计;蛋白质工程;

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专利

1. Increasing DNA substrate specificity of the EcoDam DNA-(adenine N-6)-methyltransferase by site-directed mutagenesis [J] . Elsawy H., Chahar S. Biochemistry . 2014,第11期

机译：通过定点诱变提高EcoDam DNA-（腺嘌呤N-6）-甲基转移酶的DNA底物特异性
2. Changing the DNA recognition specificity of the EcoDam DNA-(adenine-N6)-methyltransferase by directed evolution. [J] . Chahar S, Elsawy H, Ragozin Journal of Molecular Biology . 2010,第1期

机译：通过定向进化改变EcoDam DNA-（腺嘌呤-N6）-甲基转移酶的DNA识别特异性。
3. Probing the DNA Interface of the EcoRV DNA-(Adenine-N6)-methyltransferase by Site-Directed Mutagenesis, Fluorescence Spectroscopy, and UV Cross-Linking [J] . Carsten Beck, Altert Jeltsch Biochemistry . 2002,第48期

机译：通过定点诱变，荧光光谱和UV交联探测EcoRV DNA-（腺嘌呤-N6）-甲基转移酶的DNA界面
4. Effects of ligand binding on the conformation and internal dynamics in specific regions of porcine pancreatic phospholipase A{sub}2 with tryptophan as a probe: a study combining time-resolved fluorescence spectroscopy and site-directed mutagenesis [C] . Oscar Kuipers, Michel Vincent, Jean-Claude Brochon, Fluorescence Science and Technology . 2000

机译：用色氨酸作为探针的猪胰磷脂磷脂酶A {Sub} 2特定区域对特征和内部动力学的影响：一种结合时间分辨荧光光谱和定点诱变的研究
5. Regulation of Escherichia coli DNA adenine methyltransferase by non-specific interactions. [D] . Coffin, Stephanie Ruth. 2009

机译：非特异性相互作用对大肠杆菌DNA腺嘌呤甲基转移酶的调节。
6. Associations between arsenic (+3 oxidation state) methyltransferase (AS3MT) and N-6 adenine-specific DNA methyltransferase 1 (N6AMT1) polymorphisms arsenic metabolism and cancer risk in a Chilean population [O] . Rosemarie de la Rosa, Craig Steinmaus, Nicholas K Akers, -1

机译：智利人群中砷（+3氧化态）甲基转移酶（AS3MT）与N-6腺嘌呤特异性DNA甲基转移酶1（N6AMT1）多态性砷代谢和癌症风险之间的关联
7. Single amino acid changes that alter the DNA sequence specificity of the DNA-[N6-adenine] methyltransferase (Dam) of bacteriophage T4. [O] . Miner, Z, Schlagman, S L, Hattman, S 1989

机译：单个氨基酸变化会改变噬菌体T4的DNA- [N6-腺嘌呤]甲基转移酶（Dam）的DNA序列特异性。

Increasing DNA substrate specificity of the EcoDam DNA-(adenine N-6)-methyltransferase by site-directed mutagenesis

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