Orbitrap Velos with ETD for Wayne State Proteomics

描述(由申请人提供)：这项提案的目标是将一台具有ETD和纳米LC的Thermo Fisher Science Orbitrap混合FT质谱仪带到韦恩州立大学的蛋白质组学核心。该仪器具有高分辨率和1-2ppm的质量精度，具有独特的多肽测序和蛋白质鉴定能力。Orbitrap Velos的功能使其成为蛋白质组图谱、光谱计数和基于同位素的定量的特殊仪器。Orbitrap Velos的可用性和蛋白质组学核心中训练有素的操作员将是华盛顿州立大学和密歇根东南部NIH资助的研究人员研究计划的主要资产。该文书将解决目前没有得到充分满足的两个关键需求。第一种方法是基于多肽前体和子离子的准确质量进行明确的蛋白质识别。第二个是翻译后修饰(PTM)在大蛋白片段中的定位，它可以在一个多肽中维持多个翻译后修饰。目前，韦恩州立大学或底特律大都市地区唯一可用于蛋白质组分析的MS/MS仪器是具有ETD的LTQ-XL，该仪器由寻求添加Orbitrap的同一蛋白质组学核心操作。蛋白质组学设施核心是韦恩州立大学蛋白质组服务的唯一来源。大学坚定地致力于这一建议，这体现在以下支持上：1)承诺每年为核心的一般业务提供125,000美元的资金。2)购买该工具的配套资金为21.4万美元。3)从高级管理层获得75,000美元，用于支付长期服务合同的部分费用，以确保Orbitrap的持续运营。带有ETD的Orbitrap是一种最先进的系统，具有无与伦比的能力来确定多肽和蛋白质片段的准确质量，并定位蛋白质的翻译后修饰。在华盛顿州立大学，这一仪器将允许更有效地使用已经到位的蛋白质组分离仪器，并将为NIH资助的研究人员提供高效的现代蛋白质组分析途径。 公共卫生相关性：了解疾病过程和验证医学疗法需要我们发现和测量蛋白质以及细胞中蛋白质的变化。Orbitrap Velos with ETD质谱仪系统可准确识别完整蛋白质、大蛋白质片段和多肽中的蛋白质和蛋白质修饰。将Orbitrap添加到WSU蛋白质组学核心将为NIH资助的数十名研究人员提供必要的分析服务。

DESCRIPTION (provided by applicant): The goal of this proposal is to bring a Thermo Fisher Scientific Orbitrap hybrid FT mass spectrometer with ETD and nano-LC to the Proteomics Core at Wayne State University. This instrument, has unique capabilities for peptide sequencing and protein identification due to the high resolution and 1-2 ppm mass accuracy. The capabilities of the Orbitrap Velos make it an exceptional instrument fo proteomic profiling, spectral counting and isotopic based quantitation. The availability of the Orbitrap Velos and trained operators in the Proteomics Core will be a major asset to research programs of NIH-funded investigators at WSU and in the SE Michigan region. The instrument will address two critical needs that are currently not adequately met. The first of these is for unambiguous protein identification based on accurate mass of peptide precursor and daughter ions. The second is localization of post translational modifications (PTM) in large protein fragments that can maintain multiple PTMs in a single peptide. The only currently available MS/MS instruments at Wayne State University or in the metropolitan Detroit area that can be used for proteomic analysis is the LTQ-XL with ETD that is operated by the same Proteomics Core seeking to add the Orbitrap. The Proteomics Facility Core is the only source for proteomic services at Wayne State. The University is strongly committed to this proposal as evidenced by the following support: 1) A commitment to fund the general operations of the Core at $125,000 per year. 2) $214,000 in matching funds to purchase the instrument. 3) $75,000 from the SOM to defray some of the cost for a long term service contract to ensure continual operation of the Orbitrap. The Orbitrap with ETD is a state-of-the-art system with unparalleled ability to determine the accurate mass of peptides and protein fragments and to localize post-translational modifications in proteins. At WSU, this instrument will allow more productive use of instrumentation already in place for proteomic fractionation and will provide efficient access to modern proteomic analysis for NIH-funded investigators. PUBLIC HEALTH RELEVANCE: Understanding disease processes and validating medical therapies requires us to discover and measure the proteins and the changes in proteins in our cells. The Orbitrap Velos with ETD mass spectrometer system accurately identifies proteins and protein modifications in intact proteins, large protein fragments and peptides. Adding the Orbitrap to the WSU proteomics Core will provide essential analytical services to dozens of NIH funded researchers.