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S. Song, J. Liang, M. Hanson, L. T. Chylack, Jr.; Functional Role of -Crystallin in the Assembly of Lens Intermediate Filaments. Invest. Ophthalmol. Vis. Sci. 2009;50(13):2096.
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© ARVO (1962-2015); The Authors (2016-present)
The adverse effect of R120G mutation of B-crystallin on the assembly of muscle desmin intermediate filaments (IF) is well known. Recent study shows that this mutation also affects lens vimentin filaments. Since the lens contains other -crystallins (A-crystallin), we have investigated the effects of A-crystallin and its hetero-oligomers (A/B, A/R120GB, and R116CA/B) on vimentin and beaded filament structures.
We used confocal fluorescence microscopy to visualize assembly of vimentin and beaded filaments (filensin and CP49) with fusion green or red fluorescence protein (GFP or RFP) as probes. GFP-IF and -crystallin genes were cotransfected to HeLa cells and images of living cells were visualized. Protein-protein interactions were determined by FRET using either sensitized emission or photo bleaching method with the donor-acceptor pairs of GFP-IF and RFP--crystallin.
The confocal fluorescence images showed that either homo- or hetero-oligomers (A-, B-, or A/B) assist the proper assembly of IFs, but -crystallins containing myopathy-associated mutants (A/R120GB and R116CA/B) disrupt either vimentin assembly or beaded filament bundle structure. FRET analyses indicated that both A/R120GB and R116CA/B increase protein-protein interactions with both vimentin and beaded filaments.
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