Figure 5 of
Vasireddy, Mol Vis 2005;
Figure 5. FRET between wild-type and mutant ELOVL4 proteins
The physical interaction between the wild-type and mutant ELOVL4 proteins was explored by FRET. FRET measurements were made by exciting the donor at 485 nm and monitored the emission spectrum between 400-800 nm. In these experiments, green fluorescence of EGFP served as a donor (A,D,G,J), and Alexa fluor 555 conjugated V5 antibody epitope served as an acceptor (B,E,H,K). In co-transfected cells (C,F) and cells transfected with GFP-Emut and V5-E mut (L), both donors and acceptors were detected and FRET was observed. However, in cells carrying GFP-Ewt and V5-Ewt (I) no FRET was detected. Intensity was expressed as counts per second (CPS). The excitation and emission wavelengths used for EGFP and Alexa fluor labeled anti-V5-antibody are 490 nm, 509 nm, and 555 nm, 565 nm, respectively. The energy transfer observed in co-transfecetd cells indicates the close proximity between wild-type and mutant ELOVL4 (less than a 100 Å distance) and their physical interaction. These data support that the mutant and wild-type ELOVL4 proteins forms aggregates.