December 2002
Volume 43, Issue 13
Free
ARVO Annual Meeting Abstract  |   December 2002
Detection of Protein-Protein Interactions Among Lens Crystallins in a Mammalian Two-Hybrid System Assay
Author Affiliations & Notes
  • L Fu
    Ophthalmic Res/Ophthalmology Brigham and Women's Hospital/Harvard Medical School Boston MA
  • JJ Liang
    Ophthalmic Res/Ophthalmology Brigham and Women's Hospital/Harvard Medical School Boston MA
  • Footnotes
    Commercial Relationships   L. Fu, None; J.J. Liang, None. Grant Identification: NIH EY05803 and Massachusetts Lion Funds
Investigative Ophthalmology & Visual Science December 2002, Vol.43, 4798. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      L Fu, JJ Liang; Detection of Protein-Protein Interactions Among Lens Crystallins in a Mammalian Two-Hybrid System Assay . Invest. Ophthalmol. Vis. Sci. 2002;43(13):4798.

      Download citation file:


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

      ×
  • Supplements
Abstract

Abstract: : Purpose: To study protein-protein interactions among crystallins using a mammalian two-hybrid system. Methods: The major crystallin components, αA-, αB-, ß2-, and γC-crystallin genes, were subcloned into the DNA binding domain and transcription activation domain vectors of the two-hybrid system, and were co-transfected along with a reporter gene chloramphenicol acetyltransferase (CAT). Results: CAT activity indicated that there were homo- and heterogeneous interactions among αA- (or αB-), ßB2- and γC-crystallin but intensities for heterogeneous interactions of αA-ßB2, αA-γC and ßB2-γC, and homogeneous interactions of ßB2-ßB2 and γC-γC were about one-third those of αA-αA, αB-αB and αA-αB interactions. Hsp27, a member of family of the small heat-shock proteins, showed a similar interaction property with αB-crystallin. Using the N- and C-terminal domain truncated mutants, we demonstrated that both domains were important in the αA-crystallin self-interaction but that only the C-terminal domain was important in the αB-crystallin self-interaction. Additionally, we included some cataract mutant genes in the study and found changes in the interaction properties. Conclusion: These results showed that the two-hybrid system could detect interactions, both homo- and heterogeneous systems, among various crystallins.

Keywords: 378 crystallins • 527 protein structure/function • 338 cataract 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×