Abstract | PURPOSE: METHODS:
Protein- protein interactions were determined by confocal fluorescence resonance energy transfer (FRET) microscopy using green fluorescence protein (GFP) as the donor and red fluorescence protein (RFP) as the acceptor. The lens vimentin gene was fused into a GFP vector and the alphaB- crystallin (WT or R120G mutant) gene was fused into the RFP vector. The donor-acceptor plasmid pairs of intermediate filament (IF)-GFP and alphaB-RFP were co-transfected into HeLa cells. After incubation, confocal fluorescence images of the transfected cells were taken. FRET was estimated by the acceptor photobleaching method. Protein- protein interaction was evaluated by FRET efficiency. RESULTS: CONCLUSIONS: Our results show that the R120G alphaB- crystallin mutant promoted vimentin aggregation through increased protein- protein interaction. This process may contribute to the formation of congenital cataract.
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Authors | Shuhua Song, Mark J Hanson, Bing-Fen Liu, Leo T Chylack, Jack J-N Liang |
Journal | Molecular vision
(Mol Vis)
Vol. 14
Pg. 1282-7
(Jul 10 2008)
ISSN: 1090-0535 [Electronic] United States |
PMID | 18618007
(Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
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Chemical References |
- Luminescent Proteins
- Mutant Proteins
- Recombinant Fusion Proteins
- Vimentin
- alpha-Crystallin B Chain
- red fluorescent protein
- Green Fluorescent Proteins
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Topics |
- Fluorescence Resonance Energy Transfer
- Green Fluorescent Proteins
(metabolism)
- HeLa Cells
- Humans
- Lens, Crystalline
(metabolism)
- Luminescent Proteins
(metabolism)
- Microscopy, Confocal
- Mutant Proteins
(metabolism)
- Photobleaching
- Protein Binding
- Recombinant Fusion Proteins
(metabolism)
- Transfection
- Vimentin
(metabolism)
- alpha-Crystallin B Chain
(metabolism)
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