Quantification of Membrane Protein Self-Association with a High-Throughput Compatible Fluorescence Assay.
For many membrane proteins, self-association serves both structural and functional roles. Studies of such association can be simplified by switching to micelles as the membrane-mimicking environment, but native interaction is not preserved in all detergents. The selection of suitable conditions for biochemical experiments would be greatly facilitated by a quantitative high-throughput assay. Here we showed that the fluorescence polarization reduction, which resulted from homo-Förster resonance energy transfer and was measured in a high-throughput compatible format, can be used to determine both association states and constants for membrane proteins in micelles.
Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and College of Pharmaceutical Sciences, Soochow University , Suzhou, Jiangsu 215123, China.
Junbei
J
Li
Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and College of Pharmaceutical Sciences, Soochow University , Suzhou, Jiangsu 215123, China.
Xiaoyan J
0000-0003-1328-8202
XJ
Qiu
D009327:4-Chloro-7-nitrobenzofurazan / Q000737:chemistry; D020816:Amino Acid Motifs; D001622:Betaine / Q000031:analogs & derivatives / Q000737:chemistry; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D003902:Detergents; D005454:Fluorescence Polarization; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D006021:Glycophorins / Q000737:chemistry* / Q000378:metabolism; D057166:High-Throughput Screening Assays; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism; D008320:Maltose / Q000737:chemistry; D008823:Micelles; D010767:Phosphorylcholine / Q000737:chemistry; D011485:Protein Binding; D055503:Protein Multimerization; D013194:Staining and Labeling / Q000379:methods; D014763:Viral Matrix Proteins / Q000737:chemistry* / Q000378:metabolism; D000086082:Viroporin Proteins
D009327:4-Chloro-7-nitrobenzofurazan; D001622:Betaine; D003902:Detergents; D006021:Glycophorins; D008051:Lipid Bilayers; D008320:Maltose; D008823:Micelles; D010767:Phosphorylcholine; D014763:Viral Matrix Proteins; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins; C483727:sulfobetaine