RAY STEVENS PRT TIME

雷·史蒂文斯 PRT 时间

• 批准号: 8362034
• 负责人: RAYMOND C STEVENS
• 金额: $ 0.38万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8362034
• 关键词: 3-Dimensional; Active Sites; Affinity; Aldehydes; Amino Acid Substitution; Antibodies; Binding; Chorismate Mutase; Complex; DNA Sequence Rearrangement; Enzymes; Equilibrium; Funding; Grant; Immunization; National Center for Research Resources; Principal Investigator; Radiation; Reaction; Research; Research Infrastructure; Resources; Series; Source; Structure; System; United States National Institutes of Health; analog; cost; mutant; structural biology

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Immunization with transition state analogue results in a germline-encoded antibody that catalyses the rearrangement of hexadiene to aldehyde with a rate approaching that of a related pericyclic reaction catalysed by the enzyme chorismate mutase. Affinity maturation gives antibody AZ-28, which has six amino acid substitutions, one of which results in a decrease in catalytic rate. To understand the relationship between binding and catalytic rate in this system we characterized a series of active-site mutants and determined the 3-D crystal structure of the complex of AZ-28 with the transition state analogue. This analysis indicates that the activation energy depends on a complex balance of several stereoelectronic effects which are controlled by an extensive network of binding interactions in the active site.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 用过渡态类似物进行免疫会产生种系编码的抗体，该抗体催化己二烯重排为醛，其速率接近由分支酸变位酶催化的相关周环反应的速率。亲和力成熟得到抗体 AZ-28，其具有 6 个氨基酸取代，其中之一导致催化速率降低。为了了解该系统中结合和催化速率之间的关系，我们表征了一系列活性位点突变体，并确定了 AZ-28 与过渡态类似物复合物的 3-D 晶体结构。该分析表明，活化能取决于几种立体电子效应的复杂平衡，这些效应由活性位点中广泛的结合相互作用网络控制。