May 2003
Volume 44, Issue 13
ARVO Annual Meeting Abstract  |   May 2003
Effect of Oxidized ß B3-crystallin Peptide (153-167) on Thermal Aggregation of Bovine -crystallins
Author Affiliations & Notes
  • P.E. Udupa
    Department of Ophthalmology, Mason Eye Institute, University of Missouri, Columbia, MO, United States
  • K.K. Sharma
    Department of Ophthalmology and Biochemistry, Mason Eye Institute, University of Missouri, Columbia, MO, United States
  • Footnotes
    Commercial Relationships  P.E.G. Udupa, None; K.K. Sharma, None.
  • Footnotes
    Support  NIH grants EY 09855, EY11981 and unrestricted award from RPB
Investigative Ophthalmology & Visual Science May 2003, Vol.44, 2357. doi:
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      P.E. Udupa, K.K. Sharma; Effect of Oxidized ß B3-crystallin Peptide (153-167) on Thermal Aggregation of Bovine -crystallins . Invest. Ophthalmol. Vis. Sci. 2003;44(13):2357.

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

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Abstract: : Purpose: Although the mechanism for age related cataractogenesis is not clearly understood, previous studies show that there is an association between oxidation of lens crystallins, their degradation and cataractogenesis. Recently we showed that oxidized ßB3-crystallin peptide (153-167) can modulate the aggregation of denaturing ßL-crystallins (J. Biol. Chem. 277, 39136-39143). The purpose of this study was to investigate whether the same peptide would increase the thermal aggregation of bovine γ-crystallins. Method: Synthetic ßB3-crystallin peptide (153AINGTWVGYEFPGYR167) was oxidized using 0.1mM CuSO4 and 2 mM H2O2 for 3 hrs and dialyzed extensively. Thermal aggregation assay of γ-crystallin purified from bovine lens extract was performed at 55°C in 0.05M phosphate buffer containing 0.1M NaCl, pH 7.0 and the relative aggregation was measured as optical density at 360 nm. Assays were also performed with γ-crystallin with oxidized and unoxidized ßB3 peptide as well as in the presence of α-crystallin. The interaction of oxidized peptides with the aggregating γ-crystallin was also investigated using Sulfo-SBED derivatized ßB3-peptide and immuno blot studies. Results: Thermal denaturation of γ-crystallins resulted in aggregation and light scattering. Oxidized ßB3 peptide increased the aggregation of γ-crystallin compared to control with out oxidized peptide. The increase in the γ-crystallin aggregation was dependent on the concentration of the oxidized peptide. However, unoxidized ßB3 peptide in the system did not show any increase in the aggregation of γ crystallins and self-aggregation of oxidized or unoxidized peptides per see was not observed under the experimental conditions. Increased aggregation of γ-crystallin was also observed despite the presence of α-crystallin (which has anti aggregating properties) in the assay system. The Sulfo-SBED crosslinking studies confirmed the interaction between oxidized ßB3 peptide and denaturing γ-crystallin. Conclusions: These data suggest that oxidized peptides generated from ßB3-crystallin may interact with denaturing crystallins and facilitate their aggregation and light scattering, thus behaving like antichaperones. Further, this data suggests that the interaction of oxidized crystallin fragments with partially unfolded/modified crystallins may be contributing to the increased aggregation of proteins seen in aging lenses.

Keywords: oxidation/oxidative or free radical damage • crystallins • cataract 

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