Electrochemical detection of protein-protein interactions using a yeast two hybrid: 17-beta-estradiol as a model.
In this work we present a modified yeast two-hybrid bioassay for the highly sensitive detection of protein-protein interactions, based on the electrochemical monitoring of beta-D-galactosidase reporter gene activity, using p-aminophenyl-beta-D-galactopyranoside (PAPG) as a synthetic substrate. In a model system, the sensitive detection of 17-beta-estradiol was achieved at concentrations as low as 10(-11)M (approx 2 pg/ml) by monitoring 17-beta-estradiol receptor dimerization after exposure to 17-beta-estradiol. The sensitivity of this system was higher than that of standard optical methods by three orders of magnitude.
Department of Molecular Microbiology and Biotechnology, Tel-Aviv University, Ramat-Aviv 69978, Israel.
A
A
Schwartz-Mittelmann
T
T
Neufeld
D
D
Biran
J
J
Rishpon
D019281:Dimerization; D004563:Electrochemistry / Q000379:methods; D004958:Estradiol / Q000032:analysis*; D017930:Genes, Reporter; D011959:Receptors, Estradiol / Q000737:chemistry / Q000378:metabolism*; D012680:Sensitivity and Specificity; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000235:genetics*; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics
D011959:Receptors, Estradiol; D004958:Estradiol; D001616:beta-Galactosidase