Structural Studies of Metabolic Membrane Proteins

代谢膜蛋白的结构研究

• 批准号: 6611060
• 负责人: Joanne I Yeh
• 金额: $ 23.1万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6611060
• 关键词: NAD(P)H dehydrogenase; X ray crystallography; bacterial proteins; biofilm; carbohydrate metabolism; glycerol; glycerol 3 phosphate dehydrogenase; glycerol kinase; membrane proteins; microorganism metabolism; protein folding; protein protein interaction; protein structure; protein structure function; structural biology

DESCRIPTION (provided by applicant): We have obtained diffracting crystals of two membrane proteins involved in the glycerol metabolic pathway in bacteria, glycerol facilitator and glycerol-3-phosphate dehydrogenase. This is a significant step towards crystal structure determination and these structures will yield valuable new insight, linking protein folds to function and protein-protein interactions as well as mechanisms of catalysis, regulation, and transmembrane uptake of solute molecules. Underlying effective mechanisms of bacterial proliferation are means of mediating oxidative and carbohydrate metabolism. This structural study focuses on elucidating structure-function relationships of key bacterial membrane proteins that mediate fundamental oxidative and carbohydrate metabolism. We have been engaged in structural studies of these metabolic proteins for several years and are at pivotal point in integrating all of our results into a comprehensive and coherent picture of metabolism in these pathogenic bacteria. We have determined the structures of three other members of the oxidative and glycerol metabolism pathways, including NADH peroxidase, NADH oxidase, and glycerol kinase. This proposal addresses key questions correlating structure to function and regulation. Gly-3-phosphate dehydrogenase is of particular medical importance as it is a key player in providing triose phosphate intermediates for biosynthesis of polysaccharides that form biofilm and protects the bacterium from dehydration as well as antibiotic therapy. This membrane-protein structural study has the potential to yield new and novel results, which are not provided by high-throughput structural genomics. We have overcome some of the most difficult and rate-limiting hurdles in membrane protein structural studies by obtained diffracting crystals of the glycerol facilitator (2.4 A) from Gram-positive Streptococcus pneumonia, a membrane protein involved in glycerol uptake and likely to be regulated via protein-protein interactions with glycerol kinase, whose structure we've recently determined. Subtle but significant differences exist between this and the Gram-negative E. coli facilitator. We have diffracting crystals of the glycerol-3-phosphate dehydrogenase (3.3 A) from Pseudomonas aeruginosa and have promising indications for better diffracting crystals. Our specific aims are to obtain atomic resolution structures of both membrane proteins, both of which are likely to be novel targets for antibiotic design and therapy.

描述（由申请人提供）：我们已经获得了涉及细菌，甘油促剂和甘油-3-磷酸盐脱氢酶的两种膜蛋白的衍射晶体。这是朝着晶体结构确定的重要一步，这些结构将产生有价值的新见解，将蛋白质折叠与功能和蛋白质 - 蛋白质相互作用以及催化，调节和跨膜溶液分子的吸收的机制联系起来。细菌增殖的潜在有效机制是介导氧化和碳水化合物代谢的手段。这项结构研究的重点是阐明介导基本氧化和碳水化合物代谢的关键细菌膜蛋白的结构 - 功能关系。我们从事这些代谢蛋白的结构研究已有几年了，并且在这些致病细菌中代谢的全面和连贯的新陈代谢图中处于关键点。我们已经确定了其他三个氧化和甘油代谢途径的结构，包括NADH过氧化物酶，NADH氧化酶和甘油激酶。该建议解决了将结构与功能和调节相关联的关键问题。 Gly-3-磷酸脱氢酶特别重要，因为它是为多糖的生物合成提供三糖中间体的关键参与者，该多糖形成生物膜并保护细菌免受脱水和抗生素治疗。这项膜 - 蛋白质结构研究有可能产生新的和新的结果，而高通量结构基因组学并未提供。我们通过从革兰氏阳性肺炎链球菌（膜链球菌）中获得了甘油促进剂（2.4 a）的衍射晶体，从而克服了膜蛋白结构研究中一些最困难和限制的障碍，膜链球菌，膜蛋白，一种涉及甘油蛋白的膜蛋白，与甘油蛋白的摄取有关，并可能通过蛋白质蛋白质进行了蛋白质的相互作用来调节蛋白质的结构。这与革兰氏阴性大肠杆菌的促进剂之间存在细微但显着的差异。我们具有铜绿假单胞菌的甘油-3-磷酸脱氢酶（3.3 A）的衍射晶体，并具有更好的衍射晶体的有希望的适应症。我们的具体目的是获得两种膜蛋白的原子分辨率结构，这两种结构都可能是抗生素设计和治疗的新靶标。