Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes

Signe Mathiasen, Sune M. Christensen, Juan José Fung, Søren G F Rasmussen, Jonathan F. Fay, Sune K. Jorgensen, Salome Veshaguri, David L. Farrens, Maria Kiskowski, Brian Kobilka, Dimitrios Stamou

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39 Citations (Scopus)

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 b2-adrenergic receptor using ~109-fold less protein than conventional assays.

Original languageEnglish
Pages (from-to)931-934
Number of pages4
JournalNature Methods
Volume11
Issue number9
DOIs
Publication statusPublished - 3 Aug 2014
Externally publishedYes

Keywords

  • biophysical methods
  • G protein-coupled receptors
  • high-throughput screening
  • nanoscale biophysics

Cite this

Mathiasen, S., Christensen, S. M., Fung, J. J., Rasmussen, S. G. F., Fay, J. F., Jorgensen, S. K., Veshaguri, S., Farrens, D. L., Kiskowski, M., Kobilka, B., & Stamou, D. (2014). Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes. Nature Methods, 11(9), 931-934. https://doi.org/10.1038/nmeth.3062