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 Farrens, Maria Kiskowski, Brian Kobilka, Dimitrios Stamou

Research output: Contribution to journalArticle

37 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 (US)
Pages (from-to)931-934
Number of pages4
JournalNature Methods
Volume11
Issue number9
DOIs
StatePublished - 2014

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Assays
Oligomerization
Fluorescence microscopy
Fluorescence Microscopy
Adrenergic Receptors
Screening
Membrane Proteins
Proteins
Membranes
proteoliposomes

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology
  • Biochemistry
  • Cell Biology
  • Medicine(all)

Cite this

Mathiasen, S., Christensen, S. M., Fung, J. J., Rasmussen, S. G. F., Fay, J. F., Jorgensen, S. K., ... 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

Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes. / Mathiasen, Signe; Christensen, Sune M.; Fung, Juan José; Rasmussen, Søren G F; Fay, Jonathan F.; Jorgensen, Sune K.; Veshaguri, Salome; Farrens, David; Kiskowski, Maria; Kobilka, Brian; Stamou, Dimitrios.

In: Nature Methods, Vol. 11, No. 9, 2014, p. 931-934.

Research output: Contribution to journalArticle

Mathiasen, S, Christensen, SM, Fung, JJ, Rasmussen, SGF, Fay, JF, Jorgensen, SK, Veshaguri, S, Farrens, D, Kiskowski, M, Kobilka, B & Stamou, D 2014, 'Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes', Nature Methods, vol. 11, no. 9, pp. 931-934. https://doi.org/10.1038/nmeth.3062
Mathiasen S, Christensen SM, Fung JJ, Rasmussen SGF, Fay JF, Jorgensen SK et al. Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes. Nature Methods. 2014;11(9):931-934. https://doi.org/10.1038/nmeth.3062
Mathiasen, Signe ; Christensen, Sune M. ; Fung, Juan José ; Rasmussen, Søren G F ; Fay, Jonathan F. ; Jorgensen, Sune K. ; Veshaguri, Salome ; Farrens, David ; Kiskowski, Maria ; Kobilka, Brian ; Stamou, Dimitrios. / Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes. In: Nature Methods. 2014 ; Vol. 11, No. 9. pp. 931-934.
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