Proteomics Core

介绍、核心目标、进展总结以及与JHUSOM质谱的关系 核心霍普金斯DDBTRCC的蛋白质组学核心A是约翰霍普金斯医学院更大的机构质谱和蛋白质组学设施的一部分。该设施是一个国家的最先进的核心与高度复杂的，迅速发展，昂贵的设备和方法，提供给所有霍普金斯研究人员由于机构投资远远超过了一个单一的研究人员可以负担得起的设备或必要的时间，以获得专门知识的领域，只是偶尔在他们的研究。Core通过柱色谱或凝胶电泳将质谱与一维（1D）和二维（2D）分离相结合，以识别、定量或表征复杂蛋白质提取物、组分、凝胶带或斑点中的蛋白质。DDBTRCC调查人员可以获得所有设施的专业知识和服务。 霍普金斯DDBTRCC的蛋白质组学核心A与JHUSOM Mass之间的合作关系 Spectrometry Core的设计目标是使中心研究人员能够快速解析和识别复杂混合物中的目标蛋白质，以及识别作为蛋白质-蛋白质相互作用一部分的单一蛋白质。该核心的具体目标是帮助中心研究者识别在小肠、结肠、肝脏和胰腺中正常或在这些器官的疾病中差异表达的蛋白质，跟踪信号转导变化期间与结合伴侣的相互作用，识别蛋白水解切割位点并绘制翻译后修饰图。这些目标已经令人钦佩地实现，中心研究人员在这些领域中的每一个以及在发现项目中取得了成功，其中已经鉴定了GI疾病中表达改变的蛋白质。 这种合作关系使中心研究人员能够以比平常更快的周转速度和更低的成本获得我们所有的质谱和蛋白质组学服务（参见收费方法）。此外，重要的是，DDBTRC中心有一名技术专家，他的工作是了解我们的研究人员正在努力实现的目标，帮助计划实验策略，设计样品制备方法，审查生成的数据，建议对结果进行生物信息学分析的方法，以帮助指导需要验证的领域，并通过蛋白质组学核心提供的正式课程协助培训博士后研究员和学生。这种优惠待遇是DDBTRC中心非常重要的一个方面，因为它不仅加快了样品处理的速度，更重要的是增加了 成功的数据生成的概率，大大有助于确定验证数据集的方法，并有助于使用这些信息来推动调查人员的项目。 JHUSOM质谱设施由生物化学系通过科学咨询委员会管理，并由医学院院长JHUSOM基础科学部门基金的年度投入支持。基础科学主任监督所有核心，并建议或要求研究副院长为该核心的业务需求提供资金。霍普金斯DDBTRCC是通过核心申请筹集资金以部分支持JHUSOM质谱设施的四个团体之一。其他的是：国际健康营养蛋白质组学中心盖茨基金会资助，癌症中心NCI支持资助，新型治疗/艾滋病毒相关认知障碍NIMH Gran（图1）。所有赠款/合同主要调查员直接与核心主任和咨询委员会互动。 重要的是，DDBTRC中心参与JHUSOM质谱设施， 鼓励了膜蛋白处理技术的快速发展，这是大多数中心研究人员的兴趣所在。这表明了我们的核心中心成为这一安排的一部分的重要性。这是鼓励中心研究人员使用此资源的方式之一。每个支持中心（在我们的案例中为DDBTRCC）向科尔博士阐明其研究者的需求，由他决定开发/提供的服务。在近6.5年的互动中，我们的迷你中心核心PI与JHUSOM质谱设施和核心A主任R科尔合作，没有任何分歧。迷你中心的需求已经得到满足，我们中心需要的服务已经得到考虑和满足。这是一个非常积极和令人满意的关系。如果核心中心和R.科尔，出现，一个机制是在其中的核心中心PI可以解决JHUSOM质谱和蛋白质组学中心顾问委员会的分歧。 但是，预计不需要/使用这一机制，因为目前正在审议的核心活动/服务中没有分歧问题。

Introduction, Goals of Core, Summary of Progress, and Relationship to JHUSOM Mass Spectrometry Core. The Proteomics Core A of the Hopkins DDBTRCC is a part of the larger institutional Mass Spectrometryand Proteomics Facility at Johns Hopkins School of Medicine. This facility is a state-of-the-art Core with highly sophisticated, rapidly evolving, expensive equipment and approaches, made available to all Hopkins investigators due to institutional investments that far exceed what a single investigator could afford in either equipment or the time necessary to gain expertise in areas that are used only sporadically in their studies. The Core couples mass spectrometry to one (1D) and two (2D) dimensional separations by column chromatography or gel electrophoresis to identify, quantify or characterize proteins in complex protein extracts, fractions, gel bands or spots. DDBTRCC investigators have access to all facility expertise and services. The partnership between the Proteomics Core A of the Hopkins DDBTRCC and the JHUSOM Mass Spectrometry Core was designed with the goal of making available to Center Investigators the ability to rapidly resolve and identify proteins of interest from complex mixtures, as well as identifying single proteins as part of protein-protein interactions. The specific goals of this Core are to help Center Investigators identify proteins that are differentially expressed in small intestine, colon, liver and pancreas normally or in diseases of these organs, track interactions with binding partners during changes in signal transduction, identify proteolytic cleavage sites and map post-translational modifications. These goals have been admirably met, with Center Investigators succeeding in each of these areas as well as in discovery projects in which proteins altered in expression in Gl diseases have been identified. This partnership provides Center Investigators access to all our mass spectrometry and proteomics services at a more rapid turnaround than usual and at a reduced cost (sees method of charge-back). Plus, importantly, there is a technical expert assigned to the DDBTRC Center whose job is to understand what our Investigators are trying to achieve, to help plan experimental strategies, design methods of sample preparation, review the generated data, advise on approaches for bioinformatic analysis of the results to help direct areas in need of validation, and assist in training postdoctoral fellows and students via the formal courses offered by the Proteomics Core. This preferential treatment has been a very important aspect of the DDBTRC Center, because it not only has accelerated access of sample handling but more importantly has increased the probability of successful data generation, assisted greatly in determination of ways to validate the data sets, and aided in use ofthe information to move the Investigators' projects ahead. The JHUSOM Mass Spectrometry Facility is administered by the Department of Biological Chemistry via a scientific advisory board and is supported by yearly input from the JHUSOM Basic Science Departments fund from the Dean of the School of Medicine. The Basic Science Directors oversee all cores and advise or request funds from the Vice Dean of Research for operational needs of this core. The Hopkins DDBTRCC is one of four groups that has raised funds via Core applications to partially support the JHUSOM Mass Spectrometry Facility. The others are: International Health Nutriproteomics Center Gates Foundation Grant, Cancer Center NCI Support Grant, Novel Therapeutics/HIV-Associated Cognitive Disorders NIMH Gran (Fig. 1). All Grant/Contract principal investigators interact directly with the Core Director and the Advisory Board. Importantly, the DDBTRC Center involvement in the JHUSOM Mass Spectrometry Facility has encouraged rapid development of techniques to deal with membrane proteins, which is the interest of most Center Investigators. This is an indication of the importance for our Core Center of being part of this arrangement. This is one ofthe ways Center Investigators have been encouraged to use this resource. Each supporting Center (in our case DDBTRCC) clarifies the needs of its investigators to Dr. Cole who decides on services to be developed/provided. In almost 6.5 years of interactions, our Mini Center Core PI has worked with the JHUSOM Mass Spectroscopy Facility and Core A Director, R Cole with no disagreements. Needs of the Mini Center have been met and services needed by our Center have been considered and satisfied. This has been a remarkably positive and satisfying relationship. Should any issues that cannot be resolved by interactions between the Core Center and R. Cole, arise, a mechanism is in place in which the Core Center PI can address the JHUSOM Mass Spectroscopy and Proteomics Center Advisory Board over the disagreement. However, it is not anticipated that this mechanism will be required/used as there are no issues of disagreement in Core activities/services that are currently being considered.