人类 T 细胞中半胱氨酸反应性和配体结合能力的多重蛋白质组学分析。
Multiplexed proteomic profiling of cysteine reactivity and ligandability in human T cells.
机构信息
Department of Chemistry, The Scripps Research Institute, La Jolla, CA 92037, USA.
Laboratory of Chemical Immunology and Proteomics, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.
出版信息
STAR Protoc. 2021 Apr 8;2(2):100458. doi: 10.1016/j.xpro.2021.100458. eCollection 2021 Jun 18.
Differential amino acid reactivity with chemical probes can provide valuable information on the functionality and ligandability of proteins in native biological systems. Here, we present a quantitative, multiplexed chemical proteomic protocol for in-depth reactivity and ligandability profiling of cysteines in proteins in quiescent and stimulated T cells. This protocol illuminates dynamic immune state-dependent alterations in cysteine reactivity, revealing chemoselective and stereoselective small-molecule interactions with cysteines in structurally and functionally diverse proteins that lack chemical probes. For complete details on the use and execution of this protocol, please refer to Vinogradova et al. (2020).
与化学探针的差异氨基酸反应性可以为天然生物系统中蛋白质的功能和配体结合能力提供有价值的信息。在这里，我们提出了一种定量的、多重的化学蛋白质组学方法，用于深入研究静止和刺激的 T 细胞中蛋白质半胱氨酸的反应性和配体结合能力。该方案阐明了动态免疫状态依赖性半胱氨酸反应性改变，揭示了结构和功能多样化的蛋白质中半胱氨酸的化学选择性和立体选择性小分子相互作用，而这些蛋白质缺乏化学探针。有关此协议的使用和执行的完整详细信息，请参阅 Vinogradova 等人。（2020 年）。
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