Relating surfactant properties to activity and solubilization of the human adenosine a3 receptor.
The effects of various surfactants on the activity and stability of the human adenosine A3 receptor (A3) were investigated. The receptor was expressed using stably transfected HEK293 cells at a concentration of 44 pmol functional receptor per milligram membrane protein and purified using over 50 different nonionic surfactants. A strong correlation was observed between a surfactant's ability to remove A3 from the membrane and the ability of the surfactant to remove A3 selectively relative to other membrane proteins. The activity of A3 once purified also correlates well with the selectivity of the surfactant used. The effects of varying the surfactant were much stronger than those achieved by including A3 ligands in the purification scheme. Notably, all surfactants that gave high efficiency, selectivity and activity fall within a narrow range of hydrophile-lipophile balance values. This effect may reflect the ability of the surfactant to pack effectively at the hydrophobic transmembrane interface. These findings emphasize the importance of identifying appropriate surfactants for a particular membrane protein, and offer promise for the development of rapid, efficient, and systematic methods to facilitate membrane protein purification.
Berger
Bryan W
BW
Department of Chemical Engineering, University of Delaware, Newark, Delaware 19716, USA.
García
Roxana Y
RY
Lenhoff
Abraham M
AM
Kaler
Eric W
EW
Robinson
Clifford R
CR
D001667:Binding, Competitive; D001703:Biophysics / Q000379:methods*; D002244:Carbon / Q000737:chemistry; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002845:Chromatography; D003001:Cloning, Molecular; D003902:Detergents / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D004492:Edetic Acid / Q000737:chemistry; D006801:Humans; D007477:Ions; D007700:Kinetics; D008024:Ligands; D015233:Models, Statistical; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D043684:Receptor, Adenosine A3 / Q000737:chemistry*; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D013501:Surface-Active Agents / Q000737:chemistry*; D014162:Transfection
D003902:Detergents; D007477:Ions; D008024:Ligands; D043684:Receptor, Adenosine A3; D043562:Receptors, G-Protein-Coupled; D013501:Surface-Active Agents; D002244:Carbon; D004492:Edetic Acid