Thermodynamic analysis of protein-ligand binding interactions in complex biological mixtures using the stability of proteins from rates of oxidation.
Thermodynamic analysis of protein-ligand binding interactions in complex biological mixtures using the stability of proteins from rates of oxidation.
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DOI：
10.1038/nprot.2012.146
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发表时间：
2013-01
期刊：
影响因子：
14.8
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中科院分区：
文献类型：
--
作者：
The detection and quantitation of protein-ligand binding interactions is critical in a number of different areas of biochemical research from fundamental studies of biological processes to drug discovery efforts. Described here is a protocol that can be used to identify the protein targets of biologically relevant ligands (e.g. drugs like tamoxifen or cyclosporin A) in complex protein mixtures such as cell lysates. The protocol utilizes quantitative, bottom-up, shotgun proteomics technologies (iTRAQ) with a covalent labeling technique, termed Stability of Proteins from Rates of Oxidation (SPROX). In SPROX, the thermodynamic properties of proteins and protein-ligand complexes are assessed using the hydrogen peroxide-mediated oxidation of methionine residues as a function of the chemical denaturant (e.g. guanidine Hydrochloride or urea) concentration. The proteome-wide SPROX experiments described here enable the ligand binding properties of hundreds of proteins to be simultaneously assayed in the context of complex biological samples. The proteomic capabilities of the protocol render it amenable to detection of both the on- and off-target effects of ligand binding.
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影响因子：
4.4
作者：
DeArmond, Patrick D.;Xu, Ying;Strickland, Erin C.;Daniels, Kyle G.;Fitzgerald, Michael C.
通讯作者：
Fitzgerald, Michael C.
影响因子：
5.6
作者：
Liu PF;Kihara D;Park C
通讯作者：
Park C
影响因子：
7.4
作者：
West, Graham M.;Tang, Liangjie;Fitzgerald, Michael C.
通讯作者：
Fitzgerald, Michael C.
影响因子：
64.8
作者：
FIELDS, S;SONG, OK
通讯作者：
SONG, OK
影响因子：
7.4
作者：
West, Graham M.;Thompson, J. Will;Fitzgerald, Michael C.
通讯作者：
Fitzgerald, Michael C.