SOLUTION X-RAY SCATTERING STUDIES ON PROTEIN INTERACTION

蛋白质相互作用的 X 射线散射溶液研究

• 批准号: 7598192
• 负责人: MARC NIEBUHR
• 金额: $ 0.18万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-03-01 至 2008-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7598192
• 关键词: Cells; Class; Classification; Computer Retrieval of Information on Scientific Projects Database; Electrostatics; Funding; Glycerol; Grant; Institution; MYO5A gene; Measurement; Muramidase; Numbers; Property; Proteins; Publishing; Radiation; Range; Reporting; Research; Research Personnel; Resources; Roentgen Rays; Sampling; Sodium Chloride; Solubility; Solutions; Source; Structure; United States National Institutes of Health; improved; protein protein interaction

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. Systematic studies of the structure factor of a protein in solution by small angle scattering can extract electrostatic properties of the protein and protein-protein interaction. We conducted initial studies on the interactions of lysozyme, which has been extensively studied in the past in a number of published protein interaction studies. It is necessary to carry out small angle X-ray scattering measurements over a wide range of protein and salt concentrations in this class of studies. Dilute protein samples are used to obtain the form factor and highly concentrated solutions are needed for determining the structure factor. In the absence of salt the electrostatic interactions among protein molecules dominates while the attractive short-range potentials become detectable at high salt concentrations. Our preliminary results demonstrate that we can determine the lysozyme form factor at concentrations as low as 0.5 mg/ml at BL 4-2. We were able to overcome radiation damage problems we had previously observed with lysozyme, using the flow cell in combination with the solution dispenser. We plan on studying the effects of glycerol, an agent known to help minimize radiation damage on the protein interaction potential. Also studied will be the effects of L-Arg and L-Glu, which together have been reported to improve protein solubility.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 利用小角散射技术对蛋白质溶液的结构因子进行系统研究，可以提取蛋白质的静电性质和蛋白质-蛋白质相互作用。我们对溶菌酶的相互作用进行了初步研究，这在过去的一些已发表的蛋白质相互作用研究中已被广泛研究。在这类研究中，有必要在广泛的蛋白质和盐浓度范围内进行小角X射线散射测量。稀释的蛋白质样品用于获得形状因子，并且需要高浓度溶液来确定结构因子。在没有盐的情况下，蛋白质分子之间的静电相互作用占主导地位，而有吸引力的短程电位在高盐浓度下变得可检测。我们的初步结果表明，我们可以确定在浓度低至0.5毫克/毫升在BL 4-2的溶菌酶的形状因子。我们能够克服辐射损伤的问题，我们以前观察到的溶菌酶，使用流动池与溶液分配器相结合。我们计划研究甘油的影响，甘油是一种已知有助于减少蛋白质相互作用潜力的辐射损伤的试剂。还将研究L-Arg和L-Glu的作用，据报道，它们一起改善蛋白质溶解度。