PEPTIDE AND PROTEIN RECONSTITUTION IN BICELLES FOR PULSED ESR STUDY

用于脉冲 ESR 研究的 Bicelles 中的肽和蛋白质重组

• 批准号: 7602672
• 负责人: PETER P BORBAT
• 金额: $ 0.32万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7602672
• 关键词: Affect; Computer Retrieval of Information on Scientific Projects Database; Condition; Detergents; Funding; Grant; Heterogeneity; Institution; Lead; Lipid Bilayers; Liposomes; Membrane Proteins; Numbers; Peptides; Physiologic pulse; Proteins; Pulse taking; Research; Research Personnel; Resources; Sampling; Signal Transduction; Source; Spatial Distribution; Standards of Weights and Measures; United States National Institutes of Health; protein reconstitution; protein structure; reconstitution

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. The study of membrane protein structure by ESR is challenged by the non-uniform spatial distribution of proteins in the samples. The standard way of protein reconstitution in liposomes lead to their low average concentrations and increased local concentrations. Thus heterogeneity of spin distribution is manifested directly in dipolar signals as a non-linear baseline in the logarithmic plot with a non-linearity comparable in magnitude to often unresolved signals from spin pairs. The other problem is a tendency of membrane proteins to aggregate, which makes things worse. Using detergents helps to alleviate the problem. Although very useful in pulse ESR study, the detergents may affect protein structure, oligomerization number, and functionality. We see clearly that we cannot use detergents, to study peptides. In order to place a protein or peptides at more relevant conditions we are exploring reconstitution in bicelles, which as is the case of detergents would help the study of membrane proteins and peptides in lipid bilayers, since they would be isolated from each other and with heterogeneity effects greatly reduced.

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 蛋白质在样品中的空间分布不均匀，对ESR膜蛋白结构的研究提出了挑战。脂质体中蛋白质重组的标准方式导致其平均浓度较低，局部浓度较高。因此，自旋分布的不均一性在偶极信号中直接表现为对数曲线中的非线性基线，其非线性度在大小上与来自自旋对的通常无法分辨的信号相当。另一个问题是膜蛋白聚集的趋势，这使情况变得更糟。使用洗涤剂有助于缓解这个问题。虽然在脉冲ESR研究中非常有用，但洗涤剂可能会影响蛋白质的结构、寡聚化数量和功能。我们清楚地看到，我们不能使用洗涤剂来研究多肽。为了将蛋白质或多肽放置在更相关的条件下，我们正在探索在双胞体中进行重组，这就像洗涤剂的情况一样，将有助于研究脂双层中的膜蛋白和多肽，因为它们将彼此分离，并且异质性效应大大降低。