Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes
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Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes. / Mathiasen, Signe; Christensen, Sune M.; Fung, Juan José; Rasmussen, Søren Gøgsig Faarup; Fay, Jonathan F.; Jørgensen, Sune Klamer; Veshaguri, Salome; Farrens, David L.; Kiskowski, Maria; Kobilka, Brian; Stamou, Dimitrios.
In: Nature Methods, Vol. 11, 2014, p. 931-934.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
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 β2-adrenergic receptor using ∼10(9)-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 β2-adrenergic receptor using ∼10(9)-fold less protein than conventional assays.
U2 - 10.1038/nmeth.3062
DO - 10.1038/nmeth.3062
M3 - Journal article
C2 - 25086504
VL - 11
SP - 931
EP - 934
JO - Nature Methods
JF - Nature Methods
SN - 1548-7091
ER -
ID: 120586968