CHARACTERIZATION OF UDP-GALACTOPYRANOSE MUTASE

UDP-吡喃半乳糖变位酶的表征

• 批准号: 6204011
• 负责人: MICHELLE L SOLTERO
• 金额: $ 2.19万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6204011
• 关键词: NAD(H) phosphate; carbohydrate analog; enzyme mechanism; enzyme substrate; flavin adenine dinucleotide; isomerase; monosaccharides; ultraviolet spectrometry; uridine diphosphate

This proposal attempts to provide a method for characterizing the enzyme, UDP-galactopyranose mutase, which catalyzes the unfavorable conversion from UDP-galactopyranose to UDP- galactofuranose. UDP-galactofuranose is subsequently displayed on the cell surface of several microbes including Mycobacterim tuberculosis. It is not, however, displayed on the surface of eukaryotic organisms, making the enzyme a target for the development of antimicrobial agents. The mutase has been cloned from three different bacterial strains and shown to contain a flavin adenine dinucleotide (FAD) and possibly require NAD(P)H for activity. This proposal outlines a strategy using UV-visible spectroscopy to confirm these cofactors and determine their respective roles in the enzymatic mechanism. Substrate specificity of UDP-galactopyranose mutase will also be examined. By using substrate analogs of both UDP-galactopyranose and UDP- galactofuranose, identification of the important functional positions in binding to the enzyme can be explored. From this information, insight into the binding specificity and reaction mechanism of the mutase can manipulated and used for the rational design of inhibitors for the enzyme.

该建议试图提供一种表征酶，udp-galactopypyparnose突变酶的方法，该酶催化了从UDP-半乳吡喃糖到udp-galactofuranose的不利转化。 随后将UDP-Galactofuranose显示在几种微生物的细胞表面上，包括分枝杆菌结核病。 然而，它并未显示在真核生物的表面上，使该酶成为抗菌剂发展的靶标。 突变酶已从三种不同的细菌菌株中克隆，并显示出包含黄素腺嘌呤二核苷酸（FAD），可能需要NAD（P）H进行活性。 该建议概述了使用紫外可见光谱的策略来确认这些辅助因子并确定它们在酶促机制中的各自作用。 还将检查UDP-半乳吡喃糖突变酶的底物特异性。通过使用UDP-半乳吡喃糖和UDP-半乳凝集素糖的底物类似物，可以探索重要的功能位置与酶结合中的重要功能位置。 从这些信息中，可以洞悉突变酶的结合特异性和反应机制，可以操纵并用于用于酶的抑制剂的合理设计。