Sensitive detection of p65 homodimers using red-shifted and fluorescent protein-based FRET couples.

ABSTRACT:

Background

Fluorescence Resonance Energy Transfer (FRET) between the green fluorescent protein (GFP) variants CFP and YFP is widely used for the detection of protein-protein interactions. Nowadays, several monomeric red-shifted fluorescent proteins are available that potentially improve the efficiency of FRET.

Methodology/principal findings

To allow side-by-side comparison of several fluorescent protein combinations for detection of FRET, yellow or orange fluorescent proteins were directly fused to red fluorescent proteins. FRET from yellow fluorescent proteins to red fluorescent proteins was detected by both FLIM and donor dequenching upon acceptor photobleaching, showing that mCherry and mStrawberry were more efficient acceptors than mRFP1. Circular permutated yellow fluor

SUBMITTER: Goedhart J 

PROVIDER: S-EPMC1995760 | biostudies-literature | 2007 Oct

REPOSITORIES: biostudies-literature