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Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes.

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 ?2-adrenergic receptor using ?10(9)-fold less protein than conventional assays.

SUBMITTER: Mathiasen S

PROVIDER: S-EPMC4485457 | biostudies-literature | 2014 Sep

REPOSITORIES: biostudies-literature

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Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes.

Mathiasen Signe S Christensen Sune M SM Fung Juan José JJ Rasmussen Søren G F SG Fay Jonathan F JF Jorgensen Sune K SK Veshaguri Salome S Farrens David L DL Kiskowski Maria M Kobilka Brian B Stamou Dimitrios D

Nature methods 20140803 9

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 oligomeri ...[more]

PMID: 25086504

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