Using QD-FRET based method to investigate protein-protein interactions

Abstract

Förster resonance energy transfer (FRET) is an energy transfer process between a pair of light-sensitive molecules, where the donor fluorophore (initially in its electronic excited state) transfers energy to an acceptor chromophore. In this study, FRET was used to investigate the dimerization of metallothionein isoform MT-1A. The FRET system was developed based on a fluorescent quantum dots (QDs) and cyanine dye 3.5 (Cy3.5) as a versatile tool to probe small distance changes between acceptor and donor fluorophores in nanometer range. Herein, the water-soluble 450-nm emitting QDs as the donor and 590-nm Cy3.5 as the acceptor were covalently conjugated with MT-1 according to the protocol. Previous studies suggest that MTs may form oligomers under certain conditions. Therefore, further studies of this phenomenon, which has been studied here using capillary electrophoresis (CE) coupled with fluorescence detection.Förster resonance energy transfer (FRET) is an energy transfer process between a pair of light-sensitive molecules, where the donor fluorophore (initially in its electronic excited state) transfers energy to an acceptor chromophore. In this study, FRET was used to investigate the dimerization of metallothionein isoform MT-1A. The FRET system was developed based on a fluorescent quantum dots (QDs) and cyanine dye 3.5 (Cy3.5) as a versatile tool to probe small distance changes between acceptor and donor fluorophores in nanometer range. Herein, the water-soluble 450-nm emitting QDs as the donor and 590-nm Cy3.5 as the acceptor were covalently conjugated with MT-1 according to the protocol. Previous studies suggest that MTs may form oligomers under certain conditions. Therefore, further studies of this phenomenon, which has been studied here using capillary electrophoresis (CE) coupled with fluorescence detection.