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Effect of Oxidized �B3-Crystallin peptide on lens �L-Crystallin: Interaction with �B2-Crystallin

Udupa, Padmanabha EG (2005) Effect of Oxidized �B3-Crystallin peptide on lens �L-Crystallin: Interaction with �B2-Crystallin. Investigative Ophthalmology & Visual Science, 46 (7). pp. 2514-2521.

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Abstract

PURPOSE. To investigate the interaction of oxidized �B3-crystallin peptide (residues 152-166) with �L-crystallin and to identify peptide-interaction sites. METHODS. Peptides were oxidized by using CuSO4 and H2O2. Aggregation and light-scattering assays of bovine �L-crystallin were conducted at 55°C and 37°C, respectively. Assays were performed in the presence of oxidized and nonoxidized �B3- crystallin peptides and in the presence of �-crystallin. Peptideinduced change in hydrophobicity was determined by bis-ANS (4,4�-dianilino-1,1� binaphthyl-5,5� disulfonic acid) binding study. Oxidized �B3-peptide binding sites were identified by sulfo-SBED (sulfosuccinimidyl-2-[6-(biotinamido)-2-{p-azidobenzamido}- hexanoamido] ethyl-1-3 dithiopropionate) labeling and mass spectrometric analysis. RESULTS. Aggregation and relative light-scattering of �L-crystallin was higher in the presence of oxidized �B3-crystallin peptide than with �L-crystallin, without oxidized peptide and with nonoxidized peptide. Enhanced aggregation was observed despite the presence of �-crystallin in the assay. Furthermore, a significant increase in aggregation and light-scattering was observed in the presence of oxidized �B3-peptide at 37°C. Bis- ANS binding to �L-crystallin treated with oxidized �B3-peptide was two to three times higher than in the controls at 37°C. The oxidized �B3-peptide preferentially interacted with �B2-crystallin. The data were confirmed by mass spectrometric analysis. CONCLUSIONS. Oxidized �B3-peptide interacts with �B2-crystallinand enhances its aggregation and precipitation. Peptideinduced aggregation and increased hydrophobicity of the lens crystallin at 37°C are relevant to crystallin aggregation in the aging lenses.

Item Type: Article
Subjects: Medicine > KMC Manipal > Biochemistry
Depositing User: KMC Manipal
Date Deposited: 03 Aug 2015 04:48
Last Modified: 03 Aug 2015 04:48
URI: http://eprints.manipal.edu/id/eprint/143721

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