LTQ-Orbitrap Velos instrument

描述(由申请人提供)：这项提案是为了购买一台高端LC-MS/MS仪器，放置在克利夫兰诊所勒纳研究所的蛋白质组学核心实验室。该提案包含来自12名NIH资助的研究人员的项目，并要求热科学LTQ-Orbitrap Velos仪器配备Eksigent Nan1D和高效液相色谱。这是一种高端LC-MS/MS系统，代表了用于蛋白质组学研究的最先进的质谱学。目前，该实验室配备了两个LC-MS/MS系统，包括已有10年历史的ThermoScience LCQ Deca和已有7年历史的ThermoScience LTQ仪器。这些仪器一直是蛋白质组学核心的基石，在过去的十年里，该核心一直是一个富有成效的实验室，不仅为勒纳研究所的研究人员提供高质量的工作，而且为大克利夫兰科学界的研究人员提供高质量的工作。这些仪器已经跟不上勒纳研究所研究人员目前对全球蛋白质组定量实验和翻译后修饰研究的要求。我们对LTQ和LTQ-Orbitrap Velos仪器进行了直接比较，观察到与LTQ-Orbitrap Velos仪器相比，LTQ-Orbitrap Velos仪器识别的蛋白质总数增加了3到4倍，质量精度增加了一个数量级(5ppm比150ppm)，分辨率几乎提高了一个数量级(50000比6000)。收购这一最先进的仪器将使蛋白质组学核心能够更全面地分析任何给定的蛋白质组，增强识别低丰度蛋白质或翻译后修饰的能力，增加无标记定量方法的准确性，更自信地识别翻译后修饰，并允许勒纳研究所的研究人员开始在定量实验中使用稳定的同位素标记。该仪器将应用于多个项目，包括鉴定蛋白酶ADAMTS5的底物(APTE LAB)，定性和定量研究人血小板细胞骨架蛋白的蛋白酶体降解(McIntrye LAB)，更全面地了解步态复合体对炎症基因表达的翻译调控(FOX LAB)，阐明PTEN错误定位导致的蛋白表达变化在乳腺癌发生中的作用(ENG LAB)，进行乙醇和同型半胱氨酸诱导肝细胞损伤的定量蛋白质组学分析(雅各布森实验室)，确定Topo IIb调节维甲酸诱导的髓样细胞分化的机制(Ganapathi LAB)和血管平滑肌细胞对抗紧张素II反应的磷酸蛋白质组学分析(Karnik Lab)。

DESCRIPTION (provided by applicant): This proposal is for the acquisition of a high-end LC-MS/MS instrument to be placed in the Proteomics Core Laboratory at the Lerner Research Institute at the Cleveland Clinic. The proposal contains projects from twelve NIH funded investigators and is asking for a Thermoscientific LTQ-Orbitrap Velos instrument equipped with an Eksigent nano1D plus HPLC. This is a high-end LC-MS/MS system that represents the state-of-the-art in mass spectrometry that is being utilized for proteomics investigations. Currently, the lab is equipped with two LC-MS/MS systems including a ten year old Thermoscientific LCQ deca and seven year old Thermoscientific LTQ instrument. These instruments have been the cornerstone of the Proteomics Core, which for the past ten years has been a productive lab providing high quality work to investigators not only at the Lerner Research Institute but the greater Cleveland Scientific community. These instruments can no longer keep up with the current requests for global proteomic quantitative experiments and post-translational modification studies from investigators at the Lerner Research Institute. We have performed a direct comparison of the LTQ and LTQ-Orbitrap Velos instruments and observed an increase by a factor of 3 to 4 in the total number of proteins identified, an increase in mass accuracy of over an order of magnitude (5 ppm vs 150 ppm), and an increase in resolution of almost an order of magnitude (50000 vs. 6000) for the LTQ-Orbitrap Velos instrument compared to the LTQ. The acquisition of this state-of-the-art instrument will allow the Proteomics Core to more completely analyze any given proteome, enhance the ability to identify low abundant proteins or post-translational modifications, increase the accuracy of label free quantitation methods, more confidently identify post-translational modifications, and allow the researchers at the Lerner Research Institute to start utilizing stable isotope labeling in quantitative experiments. This instrument will be applied to a variety of projects including the identification of substrates for the protease ADAMTS5 (Apte lab), qualitatively and quantitatively study proteasomal degradation of cytoskeletal proteins in human platelets (McIntrye lab), more completely understand the translational control of inflammatory gene expression by the GAIT complex (Fox lab), elucidate the protein expression changes induced by mislocalization of PTEN in the context of breast carcinogenesis (Eng lab), perform a quantitative proteomic analysis of ethanol and homocysteine induce hepatic cell injury (Jacobsen lab), determine the mechanism of topo IIb regulation of retinoic acid-induced differentiation of myeloid cell (Ganapathi lab), and phosphoproteomic analysis of vascular smooth muscle cell response to antiotensin II (Karnik lab).