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于每年举行 2018 年至 2022 年夏季。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 与基因组学和代谢组学方法相结合，蛋白质组学是一个支持核心 并行探测生物活动的技术，为生物活动提供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。许多人体模型 多种动物系统，特别是脊椎动物模型中都出现了疾病 例如小鼠、斑马鱼、大鼠和青蛙，以及蠕虫和苍蝇等无脊椎动物模型；和 越来越多地利用新的干细胞技术来产生强大的体外 模型与动物模型一起部署。越来越需要实施 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，部分是通过实践和实践来实现的 高度集中的培训。 这个为期两周的密集实验室和讲座课程将重点关注尖端蛋白质组学 方法和技术。学生将获得纯化和鉴定蛋白质的实践经验 使用最新的复合物和富集/表征翻译后修饰的肽 方法。在课程的蛋白质分析部分，学生将获得以下方面的实践经验： 定量蛋白质组分析方法：将教授无标记和共价同位素标记 差异分析蛋白质组变化的方法。学生将接受高敏感性、 微毛细管高压液相色谱 (nanoHPLC) 与 Nanospray-ESI 联用 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 教授如何分析和处理鸟枪法蛋白质组学数据，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肽的转变并 设置并执行 SRM/MRM 质谱分析。他们将学习处理和解释 获取数据来测量和验证各种目标蛋白质的变化量 生物样本。对于课程的所有部分，都将重点放在数据分析上。 外部讲师将讨论课程中未直接涵盖的蛋白质组学主题和方法 包括从头序列分析； 2D 凝胶分级分离和检测/定量蛋白质水平 使用 DIGE 进行更改；先进的质谱方法；统计数据;完好无损的分析 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和 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。长期目标是培训学生发现新的机会和 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 进入人类生物学和健康的系统生物学和模式生物方法。

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.