Bioorthogonal profiling of protein methylation using azido derivative of S-adenosyl-L-methionine.
Bioorthogonal profiling of protein methylation using azido derivative of S-adenosyl-L-methionine.
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DOI：
10.1021/ja2118333
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发表时间：
2012-04-04
影响因子：
15
通讯作者：
Luo, Minkui
中科院分区：
文献类型：
--
作者：
Islam, Kabirul;Bothwell, Ian;Chen, Yuling;Sengelaub, Caitlin;Wang, Rui;Deng, Haiteng;Luo, Minkui
Protein methyltransferases (PMTs) play critical roles in multiple biological processes. Because PMTs often function in vivo through forming multimeric protein complexes, dissecting their activities in the native contexts is challenging but relevant. To address such a need, we envisioned a Bioorthogonal Profiling of Protein Methylation (BPPM) technology, in which a SAM analogue cofactor can be utilized by multiple rationally-engineered PMTs to label substrates of the corresponding native PMTs. Here 4-azido-but-2-enyl derivative of S-adenosyl-L-methionine (Ab-SAM) was reported as a suitable BPPM cofactor. The resultant cofactor-enzyme pairs were implemented to label specifically the substrates of closely related PMTs (e. g. EuHMT1 and EuHMT2) in a complex cellular mixture. The BPPM approach, coupled with mass spectrometric analysis, enables the identification of the non-histone targets of EuHMT1/2. Comparison of EuHMT1/2’s methylomes indicates that the two human PMTs, though similar in terms of their primary sequences, can act on the distinct sets of nonhistone targets. Given the conserved active sites of PMTs, Ab-SAM and its use in BPPM are expected to be transferable to other PMTs for target identification.
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影响因子：
15
作者：
Lin, Q;Jiang, FY;Gray, NS
通讯作者：
Gray, NS
影响因子：
10.5
作者：
Tachibana, M;Ueda, J;Shinkai, Y
通讯作者：
Shinkai, Y
影响因子：
4
作者：
Luo, Minkui
通讯作者：
Luo, Minkui
影响因子：
15
作者：
Wang, Rui;Zheng, Weihong;Yu, Haiqiang;Deng, Haiteng;Luo, Minkui
通讯作者：
Luo, Minkui
影响因子：
14.8
作者：
Dalhoff, C;Lukinavicius, G;Weinhold, E
通讯作者：
Weinhold, E