Cold Spring Harbor Laboratory Course on Proteomics

摘要 CSHL蛋白质组学课程 拟议的冷泉港蛋白质组学实验室课程将分别举行 从2018年到2022年的夏季。蛋白质组学是系统生物学的支柱技术之一 其中成百上千种蛋白质可以同时被监测和表征。在……里面 与基因组学和代谢组学方法相结合，蛋白质组学是一种使能的核心 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不同生物学过程。许多人类的模型 疾病在各种各样的动物系统中发生，特别是在脊椎动物模型中。 例如老鼠、斑马鱼、老鼠和青蛙，以及蠕虫和苍蝇等无脊椎动物模型；以及 越来越多地利用新的干细胞技术在体外产生强大的 模型将与动物模型一起部署。越来越多的人需要实施 系统生物学方法来处理这些模型，这需要深入了解 各种“高通量”技术的挑战和陷阱，部分是通过实践和 高度集中的训练。 这门为期两周的强化实验和讲座课程将聚焦于前沿蛋白质组学。 方法和技术。学生将获得提纯和鉴定蛋白质的实践经验 复合体及其翻译后修饰肽的富集化/表征 方法：研究方法。在本课程的蛋白质图谱部分，学生将获得以下实践经验 定量蛋白质组分析方法：将教授无标记和共价同位素标记 蛋白质组差异图谱变化的方法。学生将接受高敏感度的培训， 微毛细管高压液相色谱(NanHplc)与纳米喷雾电喷雾化联用 串联质谱仪。一维和多维分离方法的耦合 质谱学将会被教授。在课程的目标蛋白质组学部分，学生们将 教授分析和处理鸟枪式蛋白质组学数据，以便准确识别和量化 选定的蛋白质：他们将接受培训，以选择和设计目标多肽的转变，并 建立并执行SRM/MRM质谱分析。他们将学习如何处理和解释 获取数据以测量和验证各种靶蛋白中不断变化的量 生物样本。对于本课程的所有部分，将重点放在数据分析上。 外部讲师将讨论本课程未直接涉及的蛋白质组学主题和方法。 包括从头测序、2D-凝胶分级和检测/定量蛋白质水平 使用DGE的变化；先进的质谱学方法；统计；完整的分析 蛋白质和天然质谱学；以及功能蛋白质组学。 本课程的总体目标是向每个学生提供基本知识和 能够进行蛋白质组学实验和正确分析所必需的实践经验 所产生的数据。长期目标是训练学生识别新的机会和 蛋白质组学方法在他们的生物学研究中的应用，并学习如何将这些 系统生物学和人类生物学与健康的模式生物体方法。

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.