Collaborative Research: Atomic Structure Determination of ADPGlucose Pyrophosphorylase

合作研究：ADPG葡萄糖焦磷酸化酶的原子结构测定

• 批准号: 0131465
• 负责人: Jill Cupp-Vickery
• 金额: $ 24.97万
• 依托单位: University of California-Irvine
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-15 至 2006-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0131465&HistoricalAwards=false
• 关键词: Collaborative; Research; Atomic; Structure; Determination

Glycogen and starch serve as major energy storage compounds for nearly all-living organisms and are an important energy component obtained fromplants. ADP-glucose pyrophosphorylase (ADPGlc Ppase) catalyzes theconversion of glucose-1-phosphate and ATP to ADP-glucose andpyrophosphate and is a key regulated step in both bacterial glycogen andplant starch biosynthesis. ADPGlc Ppase regulation is mediated by thebinding of a number of activators and inhibitors, many of which are keymetabolic intermediates. This research utilizes the ADPGlc Ppase enzymefrom the bacteria Agrobacterium tumefaciens (Ag.t.) that is regulated by fructose-6-phosphate and pyruvate. This project outlines x-ray crystal structuralstudies on Ag.t. ADPGlc Ppase to examine substrate binding, allostericeffector binding, and the mechanism of activation. Several roadblocksassociated with x-ray crystallography have already been overcome includingcrystallization and heavy atom derivitization for phase determination. Oncethe three-dimensional structure of one member of the ADPGlc Ppase proteinfamily is determined homology modeling can be used to construct reasonablestructures of the other proteins in the family. As a major control point for theproduction of renewable and biodegradable carbon sources, ADPGlc Ppaseis an attractive enzyme target for protein engineering. Understanding theregulation of the ADPGlc Ppase enzyme family is necessary for thebioengineering of the enzyme and could lead to rational protein engineeringof the enzyme to increase starch production.

糖原和淀粉是几乎所有生物体的主要能量储存化合物，也是从植物中获得的重要能量成分。 ADP-葡萄糖焦磷酸化酶 (ADPGlc Ppase) 催化葡萄糖-1-磷酸和 ATP 转化为 ADP-葡萄糖和焦磷酸，是细菌糖原和植物淀粉生物合成中的关键调节步骤。 ADPGlc Ppase 调节是通过许多激活剂和抑制剂的结合介导的，其中许多是关键的代谢中间体。本研究利用根癌农杆菌 (Ag.t.) 细菌中的 ADPGlc Ppase 酶，该酶受 6-磷酸果糖和丙酮酸调节。该项目概述了 Ag.t 的 X 射线晶体结构研究。 ADPGlc Ppase 用于检查底物结合、变构效应物结合和激活机制。与 X 射线晶体学相关的几个障碍已经被克服，包括用于相测定的结晶和重原子衍生化。一旦确定了 ADPGlc Ppase 蛋白家族的一个成员的三维结构，同源模型就可以用于构建该家族中其他蛋白的合理结构。作为可再生和可生物降解碳源生产的主要控制点，ADPGlc Ppase 是蛋白质工程中有吸引力的酶靶标。了解 ADPGlc Ppase 酶家族的调节对于该酶的生物工程是必要的，并且可以导致对该酶进行合理的蛋白质工程以增加淀粉产量。