Multiplexed proteomic profiling of cysteine reactivity and ligandability in human T cells.
Multiplexed proteomic profiling of cysteine reactivity and ligandability in human T cells.
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DOI：
10.1016/j.xpro.2021.100458
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发表时间：
2021-06-18
期刊：
影响因子：
--
通讯作者：
Cravatt BF
中科院分区：
文献类型：
--
作者：
Vinogradova EV;Cravatt BF
不同的氨基酸与化学探针的反应性可以提供关于天然生物系统中蛋白质的功能性和配体性的有价值的信息。在这里，我们提出了一个定量的，多路化学蛋白质组学协议的半胱氨酸在静止和刺激的T细胞的蛋白质中的深入反应性和配体分析。该方案阐明了半胱氨酸反应性的动态免疫状态依赖性改变，揭示了缺乏化学探针的结构和功能多样的蛋白质中与半胱氨酸的化学选择性和立体选择性小分子相互作用。有关本方案使用和执行的完整详细信息，请参阅。半胱氨酸反应性分析用于比较不同T细胞状态下的生化变化半胱氨酸配位性分析用于发现小分子-蛋白质相互作用立体异构共价探针有助于作用机制研究不同氨基酸与化学探针的反应性可以提供有关天然生物系统中蛋白质功能和配位性的有价值信息。在这里，我们提出了一个定量的，多路化学蛋白质组学协议的半胱氨酸在静止和刺激的T细胞的蛋白质中的深入反应性和配体分析。该方案阐明了半胱氨酸反应性的动态免疫状态依赖性改变，揭示了缺乏化学探针的结构和功能多样的蛋白质中与半胱氨酸的化学选择性和立体选择性小分子相互作用。
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. Cysteine reactivity profiling to compare biochemical changes across T cell states Cysteine ligandability profiling to discover small-molecule-protein interactions Stereoisomeric covalent probes facilitate mechanism-of-action studies 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.
影响因子：
21.8
作者：
Wang, Yujia;Dix, Melissa M.;Cravatt, Benjamin F.
通讯作者：
Cravatt, Benjamin F.
影响因子：
64.8
作者：
Backus KM;Correia BE;Lum KM;Forli S;Horning BD;González-Páez GE;Chatterjee S;Lanning BR;Teijaro JR;Olson AJ;Wolan DW;Cravatt BF
通讯作者：
Cravatt BF
影响因子：
64.5
作者：
Vinogradova EV;Zhang X;Remillard D;Lazar DC;Suciu RM;Wang Y;Bianco G;Yamashita Y;Crowley VM;Schafroth MA;Yokoyama M;Konrad DB;Lum KM;Simon GM;Kemper EK;Lazear MR;Yin S;Blewett MM;Dix MM;Nguyen N;Shokhirev MN;Chin EN;Lairson LL;Melillo B;Schreiber SL;Forli S;Teijaro JR;Cravatt BF
通讯作者：
Cravatt BF