Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes – University of Copenhagen

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10 September 2014

Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes

S. Mathiasen, S.M. Christensen, J.J. Fung, S.G.F. Rasmussen, J.F. Fay, S.K. Jørgensen, S. Veshaguri, D.L. Farrens, M. Kiskowski, B. Kobilka & D. Stamou
Nature Methods 11, 931-934 (2014) doi:10.1038/nmeth.3062

Abstract
Proteoliposome reconstitution is a standard method to stabilize purified transmembrane proteins in membranes for structural and functional assays. Here we quantified intrareconstitution heterogeneities in single proteoliposomes using fluorescence microscopy. Our results suggest that compositional heterogeneities can severely skew ensemble-average proteoliposome measurements but also enable ultraminiaturized high-content screens. We took advantage of this screening capability to map the oligomerization energy of the β2-adrenergic receptor using ~109-fold less protein than conventional assays.

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