LTQ-FT MS System for On-line Protein and Glycan Analysis

描述（由申请人提供）：在生物医学质谱中心内，BUSM质谱共享仪器实验室运行两台质谱仪（Thermo-Fisher LTQ-Orbitrap和Bruker Daltonics Reflex IV MALDI-TOF MS），致力于满足NIH资助的研究人员日益增长的复杂需求。我们建议购买、安装和操作Thermo-Fisher 12-Tesla LC-LTQ FTMS系统，该系统包括机器人样品处理、一维和二维纳米液相色谱、电喷雾电离、在线电子捕获解离（ECD）和红外多光子解离（IRMPD），以提供高分辨率-高质量准确度光谱的数据依赖性采集，提高灵敏度和可靠性能，目标是实现高通量、明确的鉴定和完整的后修饰蛋白质的结构测定以及复杂碳水化合物的结构解析，以支持本提案中描述的与人类健康相关的项目。将在线记录MS和MSn光谱，在ICR室中实现ECD和IRMPD解离和高质量准确度。所要求的系统将使质谱共享仪器实验室能够实现波士顿大学不存在的功能：在线nanoLC和高通量nanospray FTMS与ECD和IRMPD碎片。这种能力将提供一组NIH资助的研究人员，他们的项目主要是翻译，获得高灵敏度，高性能的蛋白质组学和糖组学分析，重点是翻译后修饰的全面表征，包括低水平磷酸化，硝化，糖基化和其他结构多样性和/或新的修饰，这是阐明复杂疾病机制的关键。提出了九个主要和四个次要的研究项目。除了一名调查人员外，所有这些调查人员都位于BUSM校园;另一名调查人员位于马萨诸塞州总医院。公共卫生相关性主要用户的临床和基础研究包括骨吸收/骨生成（Gerstenfeld）、白血病和乳腺癌（Denis，Lerner）、心血管疾病（Cohen）、镰状细胞病（Steinberg），莱姆病（Steere），传染性微生物（萨缪尔森和罗宾斯），眼睛中的伤口愈合（Trinkaus-Randall）和伴随衰老并影响药物稳定性的蛋白质重塑（奥康纳）。未成年人用户的项目涉及淋巴瘤和乳腺癌（Demierre，Seldin）以及导致淀粉样疾病和亨廷顿病的蛋白质修饰（Costello，谢尔曼）。拟议的仪器系统将加速了解这些疾病和退行性过程的机制，从而直接有助于改善人类健康。

DESCRIPTION (provided by applicant): Within the Center for Biomedical Mass Spectrometry, the BUSM Mass Spectrometry Shared Instrumentation Laboratory operates two mass spectrometers (a Thermo-Fisher LTQ-Orbitrap and a Bruker Daltonics Reflex IV MALDI-TOF MS) dedicated to meeting the increasing and complex needs of NIH-funded investigators. We propose to purchase, install and operate a Thermo-Fisher 12-Tesla LC-LTQ FTMS system incorporating robotic sample handling, 1D and 2D nano-liquid chromatography, electrospray ionization, on-line Electron Capture Dissociation (ECD) and Infrared Multiphoton Dissociation (IRMPD), to provide data-dependent acquisition of high resolution-high mass accuracy spectra, increased sensitivity and reliable performance, with the goal to achieve high throughput, unambiguous identification and complete structural determinations of post-translationally modified proteins and structure elucidation of complex carbohydrates, to support the human health-related projects described in this proposal. MS and MSn spectra will be recorded online, with ECD and IRMPD dissociation and high mass accuracy being achieved in the ICR cell. The requested system will enable the Mass Spectrometry Shared Instrumentation Laboratory to implement functionality that does not exist at Boston University: that of on-line nanoLC- and high-throughput nanospray FTMS with ECD and IRMPD fragmentation. This capability will provide a group of NIH-funded investigators, whose projects are largely translational, with access to high sensitivity, high performance proteomics and glycomics analyses that focus on full characterization of post-translational modifications, including low-level phosphorylation, nitration, glycosylation and other structurally diverse and/or novel modifications which are keys to elucidating the mechanisms of complex diseases. Nine Major and four Minor research projects are proposed. All but one of these investigators are located on the BUSM campus; the other is located at Massachusetts General Hospital. PUBLIC HEALTH RELEVANCE The clinical and basic research of the Major Users includes bone resorption/ osteogenesis (Gerstenfeld), leukemia and breast cancer (Denis, Lerner), cardiovascular disease (Cohen), sickle cell disease (Steinberg), Lyme disease (Steere), infectious organisms (Samuelson and Robbins), wound healing in the eye (Trinkaus-Randall) and protein remodeling that accompanies aging and that affects drug stability (O'Connor). Projects of the Minor Users address lymphoma and breast cancer (Demierre, Seldin) and protein modifications that result in amyloid diseases and Huntington's Disease (Costello, Sherman). The proposed instrument system will accelerate progress towards understanding the mechanisms of these diseases and degenerative processes and will thus contribute directly toward improving human health.