Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 25086504.
- Also identified by DOI 10.1038/nmeth.3062 and PMC identifier 4485457.
- No licence information is recorded for this record.
- Because redistribution is not established, this page shows the abstract only. Follow the links below for the full text.
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 β2-adrenergic receptor using ∼10(9)-fold less protein than conventional assays.
Medical subject headings
- Algorithms
- Image Interpretation, Computer-Assisted
- Microscopy, Fluorescence
- Proteolipids
- Spectrometry, Fluorescence