Structure and function of family 1 glycosyltransferases

家族 1 糖基转移酶的结构和功能

• 批准号: 7116281
• 负责人: R Michael Garavito
• 金额: $ 25.55万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7116281
• 关键词: X ray crystallography; biosynthesis; carbohydrate structure; enzyme mechanism; enzyme structure; glycolipids; glycoproteins; glycosyltransferase; hexoses; hybrid enzyme; protein structure function; site directed mutagenesis; sugar nucleotide; uridine diphosphate glucose; vancomycin

DESCRIPTION (provided by applicant): NDP-sugar glycosyltransferases are critical for the biosynthesis of all complex carbohydrates and glycoconjugates. This large and diverse class of enzymes catalyzes the transfer of saccharide units onto the target compounds to create a diverse set of macromolecules in plants, animals, and bacteria: glycolipids, lipopolysaccharides, the glycan structures in plants, glycoproteins, and glycosylated natural products of biomedical importance like antibiotics, hormones, antitumor agents, and cardiac glycosides. Initially, we intend to determine the X-ray crystal structures of 3 subfamilies of the family 1 NDP-sugar glycosyltransferases (GTFs): GTFs involved in the biosynthesis of vancomycin group antibiotics, GTFs which create sterol glucosides, and two GTFs involved in the glycosylation of diacylglycerol. Our goals are to elucidate (1) the physical basis for the recognition of NDP-sugars, (2) the physical basis for the recognition of the aglycone acceptors, and (3) the mechanism of glycosyltransfer. Understanding the structural diversity of these enzymes will enhance our understanding of glycobiology, particularly regarding the biosynthesis of glycolipids, glycosteroids, and antibiotics. A better understanding of the glycosylation of secondary metabolites will also open up new avenues for antibiotic design against pathogenic organisms and the design of other biomedically relevant compounds (e.g., cardiac glycosides or antitumor agents). Family 1 NDP-sugar glycosyltransferases show significant conservation of functionality among homologous enzymes across species as well as between analogous enzymes within a species, despite very low levels of amino acid sequence conservation (often <25% identity) in many cases. Recent research has also detected distinct elements of structural homology that are conserved between all members of this functional class. With comparative protein sequence analysis, a structural database of the family 1 enzyme could allow the modeling of any structurally unknown protein within this family. Moreover, the relatively simple bi-domain design of family 1 NDP-sugar glycosyltransferases raises the possibility of designing chimeric enzymes with novel functionalities. Mixing hybrid enzyme design with combinatorial biosynthesis could provide practical ways to produce new glycoconjugates of biomedical importance.

描述（由申请人提供）：NDP-糖糖基转移酶对于所有复杂的碳水化合物和糖缀合物的生物合成至关重要。这种大而多样化的酶催化糖含量的转移到靶化合物上，在植物，动物和细菌中创建多种大分子：糖脂，脂多糖，脂多糖，类似于植物，糖蛋白和糖基化的自然产物的植物，糖基化植物的聚糖结构剂和心脏糖苷。最初，我们打算确定家族的3个亚家族的X射线晶体结构1 NDP-糖糖基转移酶（GTFS）：与万古霉素组抗生素的生物合成有关的GTFS，GTF的GTF，GTF，GTF，产生静脉葡萄糖苷和GTFS均与GTFS中的GTFS相关。我们的目标是阐明（1）识别NDP-糖的物理基础，（2）识别aglycone受体的物理基础，以及（3）糖基转移机制。了解这些酶的结构多样性将增强我们对糖生物学的理解，尤其是关于糖脂，糖固醇和抗生素的生物合成。对二级代谢产物的糖基化的更好理解还将为针对致病生物的抗生素设计和其他生物医学相关化合物的设计（例如心脏糖苷或抗肿瘤剂）开辟新的途径。尽管在许多情况下，但在许多情况下，家族1 NDP-糖糖基转移酶在物种之间以及类似酶之间的同源酶之间表现出显着的功能保存，尽管在许多情况下，氨基酸序列的保存非常低（通常<25％的身份）。最近的研究还检测到了该功能类别的所有成员之间保守的结构同源性的不同元素。通过比较蛋白序列分析，家族1酶的结构数据库可以允许对该家族中任何结构未知蛋白进行建模。此外，家族1 NDP糖糖基转移酶的相对简单的双域设计增加了具有具有新功能的嵌合酶的可能性。将混合酶设计与组合生物合成混合设计可以提供实用的方法来产生新的生物医学重要性糖缀合物。

项目成果

The structural basis for catalytic function of GMD and RMD, two closely related enzymes from the GDP-D-rhamnose biosynthesis pathway.

• DOI: 10.1111/j.1742-4658.2009.06993.x
• 发表时间: 2009-05
• 期刊: The FEBS journal
• 作者: King JD;Poon KKH;Webb NA;Anderson EM;McNally DJ;Brisson JR;Messner P;Garavito RM;Lam JS
• 通讯作者: Lam JS

Structure determination and analysis of acyl-CoA oxidase (ACX1) from tomato.

• DOI: 10.1107/s0907444906014107
• 发表时间: 2006-06
• 期刊: Acta crystallographica. Section D, Biological crystallography
• 作者: Rachel A. Powers;C. Rife;Anthony L. Schilmiller;Gregg A. Howe;R. Michael Garavito