Label-free quantification of membrane-ligand interactions using backscattering interferometry.
Nat Biotechnol
; 29(4): 357-60, 2011 Apr.
Article
em En
| MEDLINE
| ID: mdl-21399645
ABSTRACT
Although membrane proteins are ubiquitous within all living organisms and represent the majority of drug targets, a general method for direct, label-free measurement of ligand binding to native membranes has not been reported. Here we show that backscattering interferometry (BSI) can accurately quantify ligand-receptor binding affinities in a variety of membrane environments. By detecting minute changes in the refractive index of a solution, BSI allows binding interactions of proteins with their ligands to be measured at picomolar concentrations. Equilibrium binding constants in the micromolar to picomolar range were obtained for small- and large-molecule interactions in both synthetic and cell-derived membranes without the use of labels or supporting substrates. The simple and low-cost hardware, high sensitivity and label-free nature of BSI should make it readily applicable to the study of many membrane-associated proteins of biochemical and pharmacological interest.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Interferometria
/
Proteínas de Membrana
Limite:
Humans
Idioma:
En
Ano de publicação:
2011
Tipo de documento:
Article