SOLID-STATE NMR METHODS FOR STRUCTURAL STUDIES OF PHOSPHOLIPASE C

用于磷脂酶 C 结构研究的固态核磁共振方法

• 批准号: 8364946
• 负责人: Tatyana Polenova
• 金额: $ 3.4万
• 依托单位: UNIVERSITY OF DELAWARE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8364946
• 关键词: Architecture; Bacillus thuringiensis; Centers of Research Excellence; Crystallization; Funding; Goals; Grant; Hand; Human; Knowledge; Lipids; Magic; Membrane Proteins; Methods; NMR Spectroscopy; National Center for Research Resources; Peripheral; Phosphatidylinositols; Phospholipase; Phospholipase C; Pilot Projects; Principal Investigator; Production; Proteins; Research; Research Infrastructure; Resources; Solutions; Source; Structure; Testing; United States National Institutes of Health; X-Ray Crystallography; base; cost; interfacial; research study; solid state nuclear magnetic resonance; success; two-dimensional

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. Despite the recent successes in structural characterization of membrane proteins by X-ray crystallography and solution NMR spectroscopy, these studies remain a challenge due to the inherent insolubility and difficulties in crystallization of many membrane-associated proteins. As a result, our knowledge about the architecture and function of membrane proteins remains sparse. In particular, structural changes associated with activation of peripheral membrane proteins by non-substrate lipids, which are critical for the enzymatic activity of these proteins, are not understood at the atomic level because of a lack of experimental methods to probe specific protein-lipid interactions. In this pilot project, we are developing new magic angle spinning solid-state NMR based methods to study the structure and function of peripheral membrane proteins. In particular, two- and three-dimensional experiments are established to probe lipid-protein interactions directly. These experiments are tested on a phosphatidylinositol-specific phospholipase C (PI-PLC) from Bacillus thuringiensis. With the new methods in hand, the structural basis of the interfacial activation of PI-PLC can be examined. These efforts represent the first step toward our long-term goal to elucidate the structure and the mechanism of biomedically important human phospholipases.

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