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 This subproject (SPID 0018) is no longer active.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资助的中心赠款提供的资源的许多研究子项目之一。子弹和调查员（PI）可能已经从其他NIH来源获得了主要资金，因此可以在其他清晰的条目中代表。列出的机构适用于该中心，这不一定是调查员的机构。该子标记（蜘蛛0018）不再活跃。 Engen博士获得R01赠款后，已从NM Inbre毕业。 正在研究与蛋白质导入到线粒体相关的构象变化。了解进口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）和高分辨率质谱法（MS）方法被用来观察在最初接收将要进口到线粒体中的蛋白质的蛋白质的情况下将氘掺入蛋白质中。在此报告期间，对外膜复合物（TOM20）转运蛋白的一种蛋白质用测试蛋白（DHFR）进行了插入，并进行了HX MS分析。结果表明，TOM20能够破坏测试蛋白的动力学和/或构象，也许是一种机制，旨在使进口过程更容易展开。为了表征测试蛋白与活物能力的线粒体的相互作用，开发了质谱法。结果表明线粒体的外表面与蛋白质非常非特异性结合。我们在克服这一实验困难方面没有成功，但认为将MS友好洗涤剂纳入可能是一种解决方案。