EFFECTS OF TOM20 AND TOM22 ON THE STRUCTURE OF PROTEINS

TOM20 和 TOM22 对蛋白质结构的影响

• 批准号: 7381745
• 负责人: JOHN R ENGEN
• 金额: $ 4.97万
• 依托单位: NEW MEXICO STATE UNIVERSITY LAS CRUCES
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7381745
• 关键词: EFFECTS; TOM20; TOM22; STRUCTURE; PROTEINS

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. This subproject (SPID 0018) is no longer active. Dr. Engen has graduated from the NM-INBRE after receiving an R01 grant. Conformational changes associated with protein import into mitochondria are being investigated. Understanding the individual interactions during import will increase our understanding of this fundamental cellular process. Hydrogen exchange (HX) and high-resolution mass spectrometry (MS) methods are being used to observe the incorporation of deuterium into proteins in the presence of proteins that initially receive the protein that is to be imported into mitochondria. In this reporting period, one protein from the transporter of the outer membrane complex (Tom20) was incuabted with a test protein (DHFR) and HX MS analysis were carried out. The results indicate that Tom20 is capable of disrupting the dynamics and/or conformation of test proteins, perhaps in a mechanism designed to make unfolding easier during importation. To characterize the interaction of the test protein with live, import-competent mitochondria, mass spectrometry methods were developed. The results indicate that the outer surface of mitochondria binds quite non-specifically to proteins. We were unsuccessful in overcoming this experimental difficulty but feel that inclusion of MS friendly detergents may be a solution.

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 条目中出现。列出的机构是中心的机构，不一定是研究者的机构。该子项目 (SPID 0018) 不再活动。 Engen 博士在获得 R01 资助后从 NM-INBRE 毕业。 正在研究与蛋白质输入线粒体相关的构象变化。了解导入过程中的个体相互作用将增加我们对这一基本细胞过程的理解。氢交换 (HX) 和高分辨率质谱 (MS) 方法被用来观察在最初接收要输入线粒体的蛋白质的蛋白质存在的情况下氘与蛋白质的结合。在本报告期内，将来自外膜复合物转运蛋白 (Tom20) 的一种蛋白质与测试蛋白质 (DHFR) 一起孵育，并进行 HX MS 分析。结果表明，Tom20 能够破坏测试蛋白质的动力学和/或构象，可能是通过一种旨在使输入过程中更容易展开的机制。为了表征测试蛋白与活的具有输入能力的线粒体的相互作用，开发了质谱分析方法。结果表明线粒体外表面与蛋白质的结合非常非特异性。我们未能成功克服这一实验困难，但认为加入 MS 友好的去污剂可能是一个解决方案。