Expanding the Substrate Scope of a Bacterial Nucleotidyltransferase via Allosteric Mutations.
Expanding the Substrate Scope of a Bacterial Nucleotidyltransferase via Allosteric Mutations.
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DOI：
10.1021/acsinfecdis.2c00402
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发表时间：
2022-10-14
影响因子：
5.3
通讯作者：
Lupoli, Tania J.
中科院分区：
文献类型：
--
作者：
Zheng, Maggie;Zheng, Meng;Lupoli, Tania J.
关键词：
细菌糖结合物，如细胞表面多糖和糖蛋白，在细胞相互作用和存活中起着重要作用。被称为核苷酸转移酶的酶利用糖-1-磷酸和核苷酸三磷酸（NTPs）产生核苷酸二磷酸糖（ndp -糖），这些糖是大多数糖缀合物的基础。几十年来的研究表明，一些细菌的核苷酸转移酶具有广泛的底物耐受性，可以在体外生产各种ndp糖。虽然已知这些酶受ndp糖及其片段的变构调节，但许多工作都集中在活性位点突变的影响上。在这里，我们发现核苷基转移酶RmlA的变构位点的合理突变导致底物耐受性的扩大和催化活性的提高，这可以通过四元结构和与配体相互作用的微妙变化来解释。这些观察结果将有助于为未来研究各种细菌ndp糖和下游糖缀合物的定向生物合成提供信息。作者需要提交目录（TOC）的图形条目，与手稿标题一起，应该让读者对以下内容之一有代表性的想法：手稿中讨论的关键结构，反应，方程，概念或定理等。有关TOC图形规格，请参阅该期刊的作者说明。
Bacterial glycoconjugates, such as cell surface polysaccharides and glycoproteins, play important roles in cellular interactions and survival. Enzymes called nucleotidyltransferases use sugar-1-phosphates and nucleotide triphosphates (NTPs) to produce nucleotide diphosphate sugars (NDP-sugars), which serve as building blocks for most glycoconjugates. Research spanning several decades has shown that some bacterial nucleotidyltransferases have broad substrate tolerance and can be exploited to produce a variety of NDP-sugars in vitro. While these enzymes are known to be allosterically regulated by NDP-sugars and their fragments, much work has focused on the effect of active site mutations alone. Here, we show that rational mutations in the allosteric site of the nucleotidyltransferase RmlA leads to expanded substrate tolerance and improvements in catalytic activity that can be explained by subtle changes in quaternary structure and interactions with ligands. These observations will help inform future studies on the directed biosynthesis of diverse bacterial NDP-sugars and downstream glycoconjugates. Authors are required to submit a graphic entry for the Table of Contents (TOC) that, in conjunction with the manuscript title, should give the reader a representative idea of one of the following: A key structure, reaction, equation, concept, or theorem, etc., that is discussed in the manuscript. Consult the journal’s Instructions for Authors for TOC graphic specifications.
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DOI：
10.1038/88618
发表时间：
2001-06-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
--
作者：
Barton, WA;Lesniak, J;Nikolov, DB
通讯作者：
Nikolov, DB
影响因子：
5.2
作者：
Ahmadipour, Sanaz;Pergolizzi, Giulia;Miller, Gavin J.
通讯作者：
Miller, Gavin J.
影响因子：
6.8
作者：
Giraud, MF;Naismith, JH
通讯作者：
Naismith, JH
影响因子：
2.9
作者：
Jakeman, David L.;Young, Jessica L.;Ferrieres, Vincent
通讯作者：
Ferrieres, Vincent
影响因子：
4
作者：
Beswick, Laura;Dimitriou, Eleni;Miller, Gavin J.
通讯作者：
Miller, Gavin J.