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Relating Surfactant Properties to Activity and Solubilization of the Human Adenosine A₃ Receptor

Bryan W. Berger ^*, Roxana Y. García ^*, Abraham M. Lenhoff ^*, Eric W. Kaler ^* and Clifford R. Robinson ^* 

^* Department of Chemical Engineering, University of Delaware, Newark, Delaware; and Department of Chemistry and Biochemistry, University of Delaware and Delaware Biotechnology Institute, Newark, Delaware

Correspondence: Address reprint requests to Clifford R. Robinson, Delaware Biotechnology Institute, Delaware Technology Park, 15 Innovation Way, Newark, DE 19716. Tel.: 302-831-4942; Fax: 302-831-3447; E-mail: robinson{at}dbi.udel.edu.

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.