LTQ-Orbitrap Velos instrument

LTQ-Orbitrap Velos 仪器

• 批准号: 8050741
• 负责人: Belinda Belle Willard
• 金额: $ 60万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8050741
• 关键词: 10 year old; 7 year old; Blood Platelets; Blood Vessels; Breast; Clinic; Communities; Complex; Cytoskeletal Proteins; Ethanol; Foxes; Funding; Gene Expression; Hepatocyte; High Pressure Liquid Chromatography; Homocysteine; Homocystine; Human; Inflammatory; Investigation; Label; Laboratories; Mass Spectrum Analysis; Methods; Myeloid Cells; PTEN gene; Peptide Hydrolases; Post-Translational Protein Processing; Proteins; Proteome; Proteomics; Regulation; Research Institute; Research Personnel; Resolution; Smooth Muscle Myocytes; Stable Isotope Labeling; System; Tretinoin; United States National Institutes of Health; Work; carcinogenesis; cell injury; instrument; protein expression; research study; response

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).

描述（由申请人提供）：本提案旨在收购一台高端LC-MS/MS仪器，该仪器将放置在克利夫兰诊所Lerner研究所的蛋白质组学核心实验室。该提案包含来自12个NIH资助的研究人员的项目，并要求配备Eksigent nano 1D plus HPLC的Thermoscientific LTQ-Orbitrap Velos仪器。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技术。目前，该实验室配备了两个LC-MS/MS系统，包括一个10年历史的Thermoscientific LCQ deca和一个7年历史的Thermoscientific 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，蛋白质组学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，而且还为克利夫兰科学界提供高质量的工作。这些仪器已经无法满足Lerner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，观察到鉴定的蛋白质总数增加了3到4倍，质量准确度增加了一个数量级以上（5 ppm vs 150 ppm），与LTQ相比，LTQ-Orbitrap Velos仪器的分辨率提高了近一个数量级（50000 vs. 6000）。这一最先进仪器的获得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，提高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Lerner研究所的研究人员开始在定量实验中使用稳定同位素标记。该仪器将应用于各种项目，包括蛋白酶ADAMTS 5底物的鉴定（Apte实验室），定性和定量研究人血小板中细胞骨架蛋白的蛋白酶体降解（McIntrye实验室），更全面地了解GAIT复合物（Fox实验室）对炎症基因表达的翻译控制，阐明在乳腺癌发生过程中由PTEN的错误定位诱导的蛋白表达变化，进行乙醇和同型半胱氨酸诱导的肝细胞损伤的定量蛋白质组学分析（Jacobsen实验室），确定拓扑异构酶IIb调节视黄酸诱导的髓样细胞分化的机制（Ganapathi实验室），以及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实验室）。

项目成果

14-3-3 in Thoracic Aortic Aneurysms: Identification of a Novel Autoantigen in Large Vessel Vasculitis.

• DOI: 10.1002/art.39130
• 发表时间: 2015-07
• 期刊: Arthritis & rheumatology (Hoboken, N.J.)
• 作者: Chakravarti R;Gupta K;Swain M;Willard B;Scholtz J;Svensson LG;Roselli EE;Pettersson G;Johnston DR;Soltesz EG;Yamashita M;Stuehr D;Daly TM;Hoffman GS
• 通讯作者: Hoffman GS

Molecular Pathways Associated with Sperm Biofunction Are Not Affected by the Presence of Round Cell and Leukocyte Proteins in Human Sperm Proteome.

与精子生物功能相关的分子途径不受人类精子蛋白质组中圆细胞和白细胞蛋白存在的影响。

• DOI: 10.1021/acs.jproteome.8b00829
• 发表时间: 2019
• 期刊: Journal of proteome research
• 影响因子: 4.4
• 作者: PannerSelvam,ManeshKumar;Agarwal,Ashok;Dias,TâniaR;Martins,AnaD;Baskaran,Saradha;Samanta,Luna
• 通讯作者: Samanta,Luna

Proteomic signatures of infertile men with clinical varicocele and their validation studies reveal mitochondrial dysfunction leading to infertility.

• DOI: 10.4103/1008-682x.170445
• 发表时间: 2016-03
• 期刊: Asian journal of andrology
• 影响因子: 2.9
• 作者: Agarwal A;Sharma R;Samanta L;Durairajanayagam D;Sabanegh E
• 通讯作者: Sabanegh E

Macrophage Foam Cell-Derived Extracellular Vesicles Promote Vascular Smooth Muscle Cell Migration and Adhesion.

巨噬细胞泡沫细胞衍生的细胞外囊泡促进血管平滑肌细胞迁移和粘附。

• DOI: 10.1161/jaha.116.004099
• 发表时间: 2016-10-17
• 期刊: Journal of the American Heart Association
• 影响因子: 5.4
• 作者: Niu C;Wang X;Zhao M;Cai T;Liu P;Li J;Willard B;Zu L;Zhou E;Li Y;Pan B;Yang F;Zheng L
• 通讯作者: Zheng L

Impact of precise modulation of reactive oxygen species levels on spermatozoa proteins in infertile men.

活性氧水平的精确调节对不育男性的精子蛋白的影响。

• DOI: 10.1186/1559-0275-12-4
• 发表时间: 2015
• 期刊: Clinical proteomics
• 影响因子: 3.8
• 作者: Ayaz A;Agarwal A;Sharma R;Arafa M;Elbardisi H;Cui Z
• 通讯作者: Cui Z