VERMONT IMMUNOBIOL / INFECTIOUS DISEASES CTR: CORE C: PROTEOMICS ANALYSIS

佛蒙特州免疫生物学/传染病 CTR：核心 C：蛋白质组学分析

• 批准号: 8167729
• 负责人: Dwight E Matthews
• 金额: $ 0.91万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8167729
• 关键词: Address; Appointment; Arenavirus; Award; Biology; Caspase; Centers of Research Excellence; Communicable Diseases; Computer Retrieval of Information on Scientific Projects Database; Data; Data Analyses; Detection; Disease; Doctor of Philosophy; Electrospray Ionization; Exploratory/Developmental Grant for Diagnostic Cancer Imaging; Funding; Grant; Histamine Receptor; Human Resources; Immunobiology; Individual; Institution; Ions; Laboratories; Liquid substance; Location; Manuscripts; Mass Spectrum Analysis; Measurement; Measures; Medicine; Metabolic; Methods; Microbiology; Molecular Genetics; Organism; Peptides; Phosphorylation; Phosphorylation Site; Physiological; Post-Translational Protein Processing; Postdoctoral Fellow; Protein Binding; Proteins; Proteomics; Protocols documentation; Publishing; Research; Research Personnel; Resources; Sampling; Silicon Dioxide; Site; Source; Students; T-Cell Receptor; Techniques; Toxoplasma gondii; Training; United States National Institutes of Health; Vermont; Work; design; graduate student; improved; instrument; mass spectrometer; pathogen; professor; research study; ward

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The Proteomics Core Laboratory for the COBRE has several mass spectrometers available to assist COBRE investigators and their students. There are two Thermo-Finnigan LTQ linear quadrupole ion trap liquid chromatograph - mass spectrometers (LCMS) a Thermo-Finnigan LCQ Deca XP ion trap LCMS, and a Thermo-Finnigan LTQ Orbitrap LCMS capable of high accuracy mass measurement. All instruments have electrospray ionization (ESI) as the primary inlet, and the LTQ instruments are operated in nanospray ESI with micro-LC columns made of fused silica that also function as the nanospray tip. Nanospray improves detection limits for peptide analysis into the femtomole range. The Core continued discussion with the individual COBRE investigators to relate the capabilities of the Core and how the Core could be used to address scientific problems of COBRE investigators. Bryan Balliff, Ph.D. (Assistant Professor of Biology) who has extensive training in proteomics during his postdoctoral studies has also become involved in the Core in assisting several COBRE investigators. Dr. Ballif's laboratory is particularly designed for large scale analysis of protein phosphorylation sites. The methods that his laboratory uses routinely are being incorporated into COBRE investigator sample analyses. Personnel are available in the Core and standard protocols are in place to perform digests of proteins to peptides for proteomic analysis by LCMS, but much of the effort needs to occur in designing experiments to obtain the best mass spectrometry data. Much of the effort by Drs. Matthews and Ballif has been interacting with investigators in experiment design and data analysis for protein identification and location of posttranslational modifications. Proteomics Analyses by Mass Spectrometry The following investigators submitted samples that have been measured and completed by the Core during the period 4/1/2009  3/31/2010: Investigator Appointment Department PI: Jason Botten Assistant Professor Med-Immunobiology Div. +Joe Klaus Graduate Student +Phil Eisenhauer Technician +Joanne Russo Technician PI: Ralph Budd Professor MedImmunobiology Div. +Andreas Koenig postdoctoral fellow +Cheryl Collins Technician PI: Christopher Huston Assist. Prof Medicine  Infectious Disease +Adam Sateriale Graduate Student PI: Mariana Matrajt Assist. Prof Microbiology & Molecular Genetics PI: Cory Teuscher Professor MedImmunobiology Div. PI: Gary Ward Professor Microbiology & Molecular Genetics +Jackie Leung Graduate student An important part of the Core is working with graduate student and postdoctoral trainees, exposing them to proteomics and involving them in the sample measurement and data analysis components. The projects by Mariana Matrajt and Gary Ward focus on Toxoplasma gondii. One manuscript has been prepared and submitted by Dr. Matrajt from work performed, another from Dr. Ward was published this year in PLOS Pathogens. The project by Chris Huston is looking to continue targeted global proteomic experiments he began prior to coming to UVM. Ralph Budd and Andreas Koenig similarly are seeking to find protein binding partners to caspases and atypical T-cell receptors using proteomics approaches. Jason Botten's projects focus on the identification of host proteins that associate with arenavirus proteins and was recently awarded an NIH R21 award that heavily involved these proteomic analyses. Cory Teuscher has begun work to identify proteins that interact with histamine receptors and was recently awarded an NIH R01 award in which these proteomics experiments were an important aim. A variety of different LCMS techniques have been used in these projects ranging from identification of as many proteins as possible in the organisms being studied and changes in physiologic and metabolic conditions to focused studies of identification of protein modification sites (e.g. protein phosphorylation site identification).

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 COBRE的蛋白质组学核心实验室有几个质谱仪可用于协助COBRE研究人员和他们的学生。 有两种Thermo-Finnigan LTQ线性四极离子阱液相色谱-质谱仪（LCMS），一种是Thermo-Finnigan LCQ Deca XP离子阱LCMS，另一种是Thermo-Finnigan LTQ Orbitrap LCMS，能够进行高精度质量测量。 所有仪器都具有电喷雾电离（ESI）作为主要入口，并且LTQ仪器在具有由熔融二氧化硅制成的微型LC柱的纳米喷雾ESI中操作，所述微型LC柱也用作纳米喷雾尖端。 Nanospray将肽分析的检测限提高到飞摩尔范围。 核心继续与个别COBRE调查人员讨论，以联系核心的能力以及如何使用核心来解决COBRE调查人员的科学问题。 Bryan Balliff博士（生物学助理教授）在博士后研究期间接受了广泛的蛋白质组学培训，他也参与了核心的工作，协助几位COBRE研究人员。 Ballif博士的实验室特别设计用于蛋白质磷酸化位点的大规模分析。 他的实验室常规使用的方法正在被纳入COBRE调查员样本分析。 人员可在核心和标准的协议是在适当的地方进行蛋白质的肽的蛋白质组学分析LCMS，但大部分的努力需要发生在设计实验，以获得最佳的质谱数据。 马修斯博士和Ballif博士的大部分工作都是与研究人员在实验设计和数据分析方面进行互动，以进行蛋白质鉴定和翻译后修饰的定位。 质谱法蛋白质组学分析 以下研究者提交了在2009年4月1日期间由核心测量和完成的样本  2010年3月31日： 研究者 任命 部 PI：Jason Botten 助理教授 医学免疫生物学分部 +乔·克劳斯 研究生 +菲尔·艾森豪尔 技术员 +Joanne Russo 技术员 PI：Ralph Budd 教授 MedImmunobiology Div. +安德烈亚斯·科尼格 博士后 +谢丽尔柯林斯 技术员 PI：克里斯托弗休士顿助攻。教授 医学  传染 疾病 +Adam Sateriale 研究生 PI：Mariana Matrajt 帮忙教授 微生物学& 分子遗传学 PI：Cory Teuscher 教授 MedImmunobiology Div. PI：加里沃德 教授 微生物学& 分子遗传学 +Jackie Leung 研究生 核心的一个重要组成部分是与研究生和博士后学员合作，让他们接触蛋白质组学，并让他们参与样本测量和数据分析。 Mariana Matrajt和加里沃德的项目集中在弓形虫上。Matrajt博士根据所做的工作编写并提交了一份手稿，Ward博士的另一份手稿今年发表在PLOS Pathogens上。 克里斯休士顿的项目正在寻找继续有针对性的全球蛋白质组学实验，他开始之前来UVM。 Ralph Budd和Andreas Koenig同样也在利用蛋白质组学方法寻找半胱天冬酶和非典型T细胞受体的蛋白质结合伴侣。 Jason Botten的项目专注于鉴定与沙粒病毒蛋白相关的宿主蛋白，最近获得了NIH R21奖，该奖项主要涉及这些蛋白质组学分析。 Cory Teuscher已经开始了鉴定与组胺受体相互作用的蛋白质的工作，最近被授予NIH R 01奖，其中这些蛋白质组学实验是一个重要目标。 各种不同的LCMS技术已被用于这些项目中，从鉴定被研究的生物体中尽可能多的蛋白质，以及生理和代谢条件的变化，到鉴定蛋白质修饰位点（例如蛋白质磷酸化位点鉴定）的重点研究。