Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes

Signe Mathiasen; Sune M. Christensen; Juan José Fung; Søren Gøgsig Faarup Rasmussen; Jonathan F. Fay; Sune Klamer Jørgensen; Salome Veshaguri; David L. Farrens; Maria Kiskowski; Brian Kobilka; Dimitrios Stamou

doi:10.1038/nmeth.3062

Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes

Signe Mathiasen, Sune M. Christensen, Juan José Fung, Søren Gøgsig Faarup Rasmussen, Jonathan F. Fay, Sune Klamer Jørgensen, Salome Veshaguri, David L. Farrens, Maria Kiskowski, Brian Kobilka, Dimitrios Stamou

41 Citationer (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.

Originalsprog	Engelsk
Tidsskrift	Nature Methods
Vol/bind	11
Sider (fra-til)	931-934
Antal sider	4
ISSN	1548-7091
DOI	https://doi.org/10.1038/nmeth.3062
Status	Udgivet - 2014

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10.1038/nmeth.3062

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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.",

author = "Signe Mathiasen and Christensen, {Sune M.} and Fung, {Juan Jos{\'e}} and Rasmussen, {S{\o}ren G{\o}gsig Faarup} and Fay, {Jonathan F.} and J{\o}rgensen, {Sune Klamer} and Salome Veshaguri and Farrens, {David L.} and Maria Kiskowski and Brian Kobilka and Dimitrios Stamou",

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T1 - Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes

AU - Mathiasen, Signe

AU - Christensen, Sune M.

AU - Fung, Juan José

AU - Rasmussen, Søren Gøgsig Faarup

AU - Fay, Jonathan F.

AU - Jørgensen, Sune Klamer

AU - Veshaguri, Salome

AU - Farrens, David L.

AU - Kiskowski, Maria

AU - Kobilka, Brian

AU - Stamou, Dimitrios

PY - 2014

Y1 - 2014

N2 - 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.

AB - 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.

U2 - 10.1038/nmeth.3062

DO - 10.1038/nmeth.3062

M3 - Journal article

C2 - 25086504

SN - 1548-7091

VL - 11

SP - 931

EP - 934

JO - Nature Methods

JF - Nature Methods

ER -

Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes

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