Energetics-based discovery of protein-ligand interactions on a proteomic scale.
Energetics-based discovery of protein-ligand interactions on a proteomic scale.
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DOI：
10.1016/j.jmb.2011.02.026
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发表时间：
2011-04-22
影响因子：
5.6
通讯作者：
Park C
中科院分区：
文献类型：
--
作者：
Liu PF;Kihara D;Park C
Biochemical functions of proteins in cells frequently involve interactions with various ligands. Proteomic methods for identification of proteins that interact with specific ligands such as metabolites, signaling molecules, and drugs are valuable in investigating the regulatory mechanisms of cellular metabolism, annotating proteins with unknown functions, and also elucidating pharmacological mechanisms. Here we report an energetics-based target identification method in which target proteins in a cell lysate are identified by exploiting the effect of ligand binding on their stabilities. Urea-induced unfolding of proteins in cell lysates is probed by a short pulse of proteolysis, and the effect of a ligand on the amount of remaining folded protein is monitored on a proteomic scale. As a proof of the principle, we identified proteins that interact with ATP in the E. coli proteome. Literature and database mining confirmed a majority of the identified proteins are indeed ATP-binding proteins. Four identified proteins that were previously not known to interact with ATP were cloned and expressed to validate the result. Except for one protein, the effects of ATP on urea-induced unfolding were confirmed. Analyses of the protein sequences and structure models were also employed to predict potential ATP binding sites in the identified proteins. Our results demonstrate that this energetics-based target identification approach is a facile method to identify proteins that interact with specific ligands on a proteomic scale.
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DOI：
10.1107/s1744309108029515
发表时间：
2008-10-01
影响因子：
0.9
作者：
Baba-Dikwa, Aisha;Thompson, Darren;Watson, Kimberly A.
通讯作者：
Watson, Kimberly A.
DOI：
10.1111/j.1432-1033.1973.tb03059.x
发表时间：
1973-01-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
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KOSAKOWSKI, HM;HOLLER, E
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HOLLER, E
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8
作者：
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通讯作者：
Park, Chiwook
影响因子：
14.9
作者：
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通讯作者：
Thornton JM
DOI：
10.1111/j.1432-1033.1991.tb16239.x
发表时间：
1991-09-15
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
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通讯作者：
ROY, S