LC-Tandem Mass Spectrometer

描述（由申请人提供）：了解基因组如何控制生物系统的生理需要对所产生的蛋白质进行详细分析。在过去的几年里，质谱和生物信息学方法的技术改进使分析蛋白质种群（蛋白质组学）的能力提高了一个数量级，这对俄亥俄医学院和邻近机构的研究项目至关重要。本提案的目的是获得一种液相色谱-串联质谱（LC/MS/MS）仪器，以及所需的核心配件，以使nih资助的主要用户以及来自俄亥俄医学院、托莱多大学和鲍灵格林州立大学的各种次要用户最有效地使用它。与DNA测序不同，高效的LC/MS/MS蛋白质组学分析需要研究团队和实验室核心人员之间的密切互动。在LC/MS/MS中尤其如此，其中一群色氨酸肽被分解，从门控离子阱中释放出来，通过与惰性气体原子的碰撞而破碎，并通过分析所得离子的质量指纹来测序。俄亥俄医学院已经有了容纳拟议设备的实验室空间，以及确保其最佳使用所需的专业知识。

DESCRIPTION (provided by applicant): Understanding how genomes control the physiology of biological systems requires detailed analysis of the proteins produced. In the past several years, technical refinements to mass spectroscopy and bioinformatics methods have increased the power of analyzing protein populations (proteomics) by another order of magnitude, making it vital to research projects at the Medical College of Ohio and neighboring institutions. The purpose of this proposal is to obtain a liquid chromatography - tandem mass spectroscopy (LC/MS/MS) instrument, along with the core accessories required to allow its most efficient use by NIH-funded major users, and a variety of minor users from the Medical College of Ohio, University of Toledo, and Bowling Green State University. Unlike DNA sequencing, efficient proteomic analysis by LC/MS/MS requires close interactions between the research team and core laboratory personnel. This is particularly true of LC/MS/MS, in which a population of tryptic peptides are resolved, released from a gated ion trap, fragmented by collision with atoms of an inert gas, and sequenced by analyzing of the mass fingerprint of the resulting ions. Laboratory space to house the proposed equipment, and expertise required to ensure its optimal use, are already in place at the Medical College of Ohio. Major and minor users of the proposed LC/MS/MS equipment have been identified. The NIH-funded projects of the eight major users include characterization of the fungal pathogen and potential bioterrorism agent Coccidioides immitis, the link between reactive oxygen species-dependent inactivation of Na + ATPase and pathological changes in cardiac myocytes, characterization of proteins interacting with Na + ATPase, the role of the CEACAM1 cell adhesion molecule in insulin metabolism, regulation of replication of the potential gene therapy vector adeno-associated virus (AAV), characterization of the MAP kinase inhibitory protein RKIP, investigating the role of Rab GTPases in controlling cell growth, and characterizing the endocytosis and recycling of the GPI-anchored folate receptor. All of these projects, and others as well, will be substantially strengthened by the requested equipment.