May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Confocal Fluorescence Microscopy Studies of Human Lens T5P C-Crystallin Mutant
Author Affiliations & Notes
  • J. J. Liang
    Ophthalmic Research/Surgery, Brigham & Womens Hospital/Harvard Medical School, Boston, Massachusetts
  • B.-F. Liu
    Ophthalmic Research/Surgery, Brigham & Womens Hospital/Harvard Medical School, Boston, Massachusetts
  • M. Hanson
    Ophthalmic Research/Surgery, Brigham & Womens Hospital/Harvard Medical School, Boston, Massachusetts
  • S. Song
    Ophthalmic Research/Surgery, Brigham & Womens Hospital/Harvard Medical School, Boston, Massachusetts
  • Footnotes
    Commercial Relationships  J.J. Liang, None; B. Liu, None; M. Hanson, None; S. Song, None.
  • Footnotes
    Support  NIH Grant EY13968 and Massachusetts Lions Eye Research Fund
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 4088. doi:https://doi.org/
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    • Get Citation

      J. J. Liang, B.-F. Liu, M. Hanson, S. Song; Confocal Fluorescence Microscopy Studies of Human Lens T5P C-Crystallin Mutant. Invest. Ophthalmol. Vis. Sci. 2008;49(13):4088. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : To investigate protein aggregation of T5P γC-crystallin and protein-protein interaction with αA- crystallin.

Methods: : Crystallin genes (T5PCRYGC, CRYGC, CRYAA) were fused to either green fluorescence protein (GFP) or red fluorescence protein (RFP) vector. Donor-acceptor plasmid pairs, such as T5PγC-GFP and αA-RFP, were cotransfected into HeLa cells. Confocal fluorescence images of the cultured cells were taken. Protein-protein interaction was evaluated by fluorescence resonance energy transfer (FRET) between the donor and acceptor.

Results: : The confocal fluorescence images show that T5P γC-crystallin expression contains a large amount of protein aggregates but co-expression with either γC- or αA-crystallin reduces the aggregation considerably. FRET determination indicates that T5PγC-T5PγC shows less protein-protein interaction than either T5PγC-γC or T5PγC-αA.

Conclusions: : Our results demonstrate that αA- and γC-crystallin can rescue T5P γC-crystallin from aggregation. The mechanism is through increased protein-protein interaction.

Keywords: crystallins • microscopy: light/fluorescence/immunohistochemistry • mutations 
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