Cold Spring Harbor Laboratory Course on Proteomics

摘要 蛋白质组学课程 拟议中的冷泉港蛋白质组学实验室课程将于 从2018年到2022年的夏天。蛋白质组学是系统生物学的支柱技术之一， 可以同时监测和表征数百或数千种蛋白质。在 蛋白质组学与基因组学和代谢组学方法相结合， 并行探测生物活动的技术，提供了前所未有的分析能力， 哺乳动物发育和疾病中的多种生物过程。许多人的模型 疾病已经在多种动物系统，特别是脊椎动物模型中发展 例如小鼠、斑马鱼、大鼠和青蛙，以及无脊椎动物模型例如蠕虫和苍蝇;以及 越来越多的新干细胞技术的使用正在被利用来产生强大的体外 与动物模型一起使用。越来越需要执行 系统生物学的方法，这些模型，这需要深入了解 挑战和陷阱的各种“高通量”技术，实现了部分动手， 高度集中的训练。 这个密集的两周实验室和讲座课程将集中在尖端的蛋白质组学 方法和技术。学生将获得纯化和鉴定蛋白质的实践经验 复合物和富集/表征后修饰的肽，使用最新的 方法.在本课程的蛋白质分析部分，学生将获得实践经验， 定量蛋白质组分析方法：将教授无标记和共价同位素标记 方法来差异分析蛋白质组中的变化。学生将接受高灵敏度， 微毛细管高压液相色谱（nanoHPLC）与nanospray-ESI偶联， 串联质谱法单维和多维分离方法耦合 将教授质谱分析。在课程的目标蛋白质组学部分，学生将 教授分析和处理鸟枪蛋白质组学数据，以准确地识别和定量 选定的蛋白质：他们将接受培训，以选择和设计目标肽的转换， 设置并执行SRM/MRM质谱分析。他们将学习处理和解释 获得的数据，以测量和验证各种靶向蛋白质的变化量， 生物样本对于课程的所有部分，将重点放在数据分析上。 外部讲师将讨论蛋白质组学的主题和方法没有直接涉及的课程 包括从头序列分析; 2D-凝胶分级和检测/定量蛋白质水平 DIGE变化;高级质谱法;统计学;完整的 蛋白质和天然质谱;和功能蛋白质组学。 课程的总体目标是为每个学生提供基础知识， 能够进行蛋白质组学实验和正确分析所需的实践经验 结果数据。长期目标是培养学生识别新的机会， 蛋白质组学方法在生物学研究中的应用，并学习如何整合这些方法。 系统生物学和模式生物学方法来研究人类生物学和健康。

Abstract CSHL Proteomics Course The proposed Cold Spring Harbor Laboratory Course on Proteomics is to be held each summer from 2018 through 2022. Proteomics is one of the pillar technologies of systems biology by which hundreds or thousands of proteins can be monitored and characterized simultaneously. In combination with genomics and metabolomics approaches, proteomics is an enabling core technology to probe biological activities in parallel, providing unprecedented analytical power into diverse biological processes in mammalian development and disease. Models of many human diseases have been developed in a wide variety of animal systems, in particular vertebrate models such as mouse, zebrafish, rat and frog, and invertebrate models such as worms and flies; and increasingly the use of new stem cell technologies is being harnessed to produce powerful in vitro models to be deployed alongside animal models. There is a growing need for implementation of systems biology approaches to these models, which necessitates an in-depth understanding of the challenges and pitfalls of various “high-throughput” technologies, achieved in part by hands-on and highly focused training. This intensive two week laboratory and lecture course will focus on cutting-edge proteomics approaches and technologies. Students will gain practical experience purifying and identifying protein complexes and enriching/characterizing post-translationally modified peptides using the latest methods. In the protein profiling portion of the course, students will gain hands-on experience in quantitative proteome analysis methods: they will be taught label-free and covalent isotopic-labeling approaches to differentially profile changes in proteomes. Students will be trained in high-sensitivity, microcapillary high pressure liquid chromatography (nanoHPLC) coupled with nanospray-ESI and tandem mass spectrometry. Both single dimension and multidimensional separation methods coupled to mass spectrometry will be taught. In the targeted proteomics section of the course, students will be taught to analyze and process shotgun proteomics data in order to accurately identify and quantify selected proteins: they will be trained to select and design transitions for targeted peptides and to setup and perform SRM/MRM mass spectrometry assays. They will learn to process and interpret the acquired data to measure and validate changing quantities of targeted proteins in a variety of biological samples. For all sections of the course, a strong emphasis will be placed on data analysis. Outside lecturers will discuss proteomics topics and methods not directly covered in the course including de novo sequence analysis; 2D-gel fractionations and detecting/quantifying protein-level changes using DIGE; advanced mass spectrometry methods; statistics; the analysis of intact proteins, and native mass spectrometry; and functional proteomics. The overall aim of the course is to provide each student with the fundamental knowledge and hands-on experience necessary to be able to perform proteomics experiments and properly analyze the resulting data. The long term goal is to train students to identify new opportunities and applications for proteomics approaches in their biological research and to learn how to integrate these into systems biology and model organism approaches to human biology and health.