Identification of lysine acetyltransferase p300 substrates using 4-pentynoyl-coenzyme A and bioorthogonal proteomics.
Identification of lysine acetyltransferase p300 substrates using 4-pentynoyl-coenzyme A and bioorthogonal proteomics.
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DOI：
10.1016/j.bmcl.2011.05.060
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发表时间：
2011-09-01
影响因子：
2.7
通讯作者：
Howard, Hang C.
中科院分区：
文献类型：
--
作者：
Yu-Ying, Yang;Markus, Grammel;Howard, Hang C.
蛋白质组学研究已经在真核生物和细菌中鉴定出大量的赖氨酸乙酰化蛋白。确定负责每个蛋白质乙酰化标记的单个赖氨酸乙酰转移酶对于阐明潜在的调控机制至关重要，但由于有限的生物化学方法而具有挑战性。在这里，我们描述了生物正交化学蛋白质组学方法的应用，个人赖氨酸乙酰转移酶的配置文件和识别基板。添加4-戊炔酰辅酶A，一个炔基化学报告蛋白乙酰化，细胞提取物，与纯化的赖氨酸乙酰转移酶p300，使荧光分析和鉴定蛋白质底物通过铜（I）催化的炔-叠氮化物环加成。我们确定了几个已知的蛋白质底物的乙酰转移酶p300以及赖氨酸残基的修改。有趣的是，使用这种方法还发现了几种新的候选p300底物及其乙酰化位点。我们的研究结果表明，生物正交化学蛋白质组学允许在体外快速底物识别的个别蛋白乙酰转移酶。
Proteomic studies have identified a plethora of lysine acetylated proteins in eukaryotes and bacteria. Determining the individual lysine acetyltransferases responsible for each protein acetylation mark is crucial for elucidating the underlying regulatory mechanisms, but has been challenging due to limited biochemical methods. Here, we describe the application of a bioorthogonal chemical proteomics method to profile and identify substrates of individual lysine acetyltransferases. Addition of 4-pentynoyl-coenzyme A, an alkynyl chemical reporter for protein acetylation, to cell extracts, together with purified lysine acetyltransferase p300, enabled the fluorescent profiling and identification of protein substrates via Cu(I)-catalyzed alkyne-azide cycloaddition. We identified several known protein substrates of the acetyltransferase p300 as well as the lysine residues that were modified. Interestingly, several new candidate p300 substrates and their sites of acetylation were also discovered using this approach. Our results demonstrate that bioorthogonal chemical proteomics allows the rapid substrate identification of individual protein acetyltransferases in vitro.
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影响因子：
64.5
作者：
An, W;Kim, J;Roeder, RG
通讯作者：
Roeder, RG
影响因子：
56.9
作者：
Choudhary, Chunaram;Kumar, Chanchal;Mann, Matthias
通讯作者：
Mann, Matthias
影响因子：
64.5
作者：
Lin YY;Lu JY;Zhang J;Walter W;Dang W;Wan J;Tao SC;Qian J;Zhao Y;Boeke JD;Berger SL;Zhu H
通讯作者：
Zhu H
DOI：
10.1073/pnas.51.5.786
发表时间：
1964-01-01
影响因子：
11.1
作者：
ALLFREY, VG;FAULKNER, R;MIRSKY, AE
通讯作者：
MIRSKY, AE
代谢酶的乙酰化协调碳源利用和代谢通量
DOI：
10.1126/science.1179687
发表时间：
2010-02-19
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Wang Q;Zhang Y;Yang C;Xiong H;Lin Y;Yao J;Li H;Xie L;Zhao W;Yao Y;Ning ZB;Zeng R;Xiong Y;Guan KL;Zhao S;Zhao GP
通讯作者：
Zhao GP