Quantifying the stabilizing effects of protein-ligand interactions in the gas phase

被引:129
作者
Allison, Timothy M. [1 ]
Reading, Eamonn [1 ]
Liko, Idlir [1 ]
Baldwin, Andrew J. [1 ]
Laganowsky, Arthur [1 ]
Robinson, Carol V. [1 ]
机构
[1] Univ Oxford, Dept Chem, Oxford OX1 5QY, England
来源
NATURE COMMUNICATIONS | 2015年 / 6卷
基金
英国生物技术与生命科学研究理事会;
关键词
MOBILITY-MASS-SPECTROMETRY; NEGATIVE COOPERATIVITY; MEMBRANE-PROTEINS; COMPLEXES; BINDING; DETERGENT; INFORMATION; ASSAY;
D O I
10.1038/ncomms9551
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The effects of protein-ligand interactions on protein stability are typically monitored by a number of established solution-phase assays. Few translate readily to membrane proteins. We have developed an ion-mobility mass spectrometry approach, which discerns ligand binding to both soluble and membrane proteins directly via both changes in mass and ion mobility, and assesses the effects of these interactions on protein stability through measuring resistance to unfolding. Protein unfolding is induced through collisional activation, which causes changes in protein structure and consequently gas-phase mobility. This enables detailed characterization of the ligand-binding effects on the protein with unprecedented sensitivity. Here we describe the method and software required to extract from ion mobility data the parameters that enable a quantitative analysis of individual binding events. This methodology holds great promise for investigating biologically significant interactions between membrane proteins and both drugs and lipids that are recalcitrant to characterization by other means.
引用
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页数:10
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