STUDIES OF CRYO-COOLING AND CATALYTIC MECHANISM IN E COLI BETA-GALACTOSIDASE

大肠杆菌β-半乳糖苷酶低温冷却及催化机制的研究

• 批准号: 7370342
• 负责人: Douglas Henry Juers
• 金额: $ 0.02万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7370342
• 关键词: STUDIES; CRYO; COOLING; CATALYTIC; MECHANISM

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Three experiments are described involving E. coli beta-galactosidase designed to address three broadly related topics. First is to improve our basic knowledge about cryocooling by understanding the cryocooling induced repacking in crystals of beta-galactosidase. Second is to understand how packing forces modulate enzyme activity by characterizing differences in ligand binding to separate subunits in an asymmetric unit. And third is to understand the role of a conformational switch in the activity of beta-galactosidase and how packing forces affect this switch. Each experiment is straightforward in principle, but technically demanding given the large size of the unit cell. Structures of variants and complexes will be determined up to 1.1 ¿¿resolution.

该子项目是利用 NIH/NCRR 资助的中心拨款提供的资源的众多研究子项目之一。子项目和研究者 (PI) 可能已从另一个 NIH 来源获得主要资金，因此可以在其他 CRISP 条目中得到体现。列出的机构是中心的机构，不一定是研究者的机构。描述了涉及大肠杆菌β-半乳糖苷酶的三个实验，旨在解决三个广泛相关的主题。首先是通过了解低温冷却诱导的 β-半乳糖苷酶晶体重新堆积来提高我们关于低温冷却的基础知识。 其次是了解堆积力如何通过表征配体与不对称单元中单独亚基的结合差异来调节酶活性。第三是了解构象转换在 β-半乳糖苷酶活性中的作用以及包装力如何影响这种转换。每个实验原则上都很简单，但由于晶胞尺寸较大，技术要求很高。变体和复合物的结构将以高达 1.1 的分辨率确定。