Abstract
PURPOSE. To investigate the interaction of oxidized βB3-crystallin peptide (residues 152-166) with βL-crystallin and to identify peptide-interaction sites. METHODS. Peptides were oxidized by using CuSO 4 and H2O2. Aggregation and light-scattering assays of bovine βL-crystallin were conducted at 55°C and 37°C, respectively. Assays were performed in the presence of oxidized and nonoxidized βB3-crystallin peptides and in the presence of α-crystallin. Peptide-induced change in hydrophobicity was determined by bis-ANS (4,4′-dianilino-1,1′ binaphthyl-5,5′ disulfonic acid) binding study. Oxidized βB3-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 βL-crystallin was higher in the presence of oxidized βB3-crystallin peptide than with βL-crystallin, without oxidized peptide and with nonoxidized peptide. Enhanced aggregation was observed despite the presence of α-crystallin in the assay. Furthermore, a significant increase in aggregation and light-scattering was observed in the presence of oxidized βB3-peptide at 37°C. Bis-ANS binding to βL-crystallin treated with oxidized βB3-peptide was two to three times higher than in the controls at 37°C. The oxidized βB3-peptide preferentially interacted with βB2-crystallin. The data were confirmed by mass spectrometric analysis. CONCLUSIONS. Oxidized βB3-peptide interacts with βB2-crystallin and enhances its aggregation and precipitation. Peptide-induced aggregation and increased hydrophobicity of the lens crystallin at 37°C are relevant to crystallin aggregation in the aging lenses.
| Original language | English |
|---|---|
| Pages (from-to) | 2514-2521 |
| Number of pages | 8 |
| Journal | Investigative Ophthalmology and Visual Science |
| Volume | 46 |
| Issue number | 7 |
| DOIs | |
| Publication status | Published - 01-12-2005 |
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All Science Journal Classification (ASJC) codes
- Ophthalmology
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Effect of oxidized βB3-crystallin peptide on lens βL- crystallin : Interaction with βB2-crystallin. / Udupa, E. G Padmanabha; Sharma, K. Krishna.
In: Investigative Ophthalmology and Visual Science, Vol. 46, No. 7, 01.12.2005, p. 2514-2521.Research output: Contribution to journal › Article
TY - JOUR
T1 - Effect of oxidized βB3-crystallin peptide on lens βL- crystallin
T2 - Interaction with βB2-crystallin
AU - Udupa, E. G Padmanabha
AU - Sharma, K. Krishna
PY - 2005/12/1
Y1 - 2005/12/1
N2 - PURPOSE. To investigate the interaction of oxidized βB3-crystallin peptide (residues 152-166) with βL-crystallin and to identify peptide-interaction sites. METHODS. Peptides were oxidized by using CuSO 4 and H2O2. Aggregation and light-scattering assays of bovine βL-crystallin were conducted at 55°C and 37°C, respectively. Assays were performed in the presence of oxidized and nonoxidized βB3-crystallin peptides and in the presence of α-crystallin. Peptide-induced change in hydrophobicity was determined by bis-ANS (4,4′-dianilino-1,1′ binaphthyl-5,5′ disulfonic acid) binding study. Oxidized βB3-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 βL-crystallin was higher in the presence of oxidized βB3-crystallin peptide than with βL-crystallin, without oxidized peptide and with nonoxidized peptide. Enhanced aggregation was observed despite the presence of α-crystallin in the assay. Furthermore, a significant increase in aggregation and light-scattering was observed in the presence of oxidized βB3-peptide at 37°C. Bis-ANS binding to βL-crystallin treated with oxidized βB3-peptide was two to three times higher than in the controls at 37°C. The oxidized βB3-peptide preferentially interacted with βB2-crystallin. The data were confirmed by mass spectrometric analysis. CONCLUSIONS. Oxidized βB3-peptide interacts with βB2-crystallin and enhances its aggregation and precipitation. Peptide-induced aggregation and increased hydrophobicity of the lens crystallin at 37°C are relevant to crystallin aggregation in the aging lenses.
AB - PURPOSE. To investigate the interaction of oxidized βB3-crystallin peptide (residues 152-166) with βL-crystallin and to identify peptide-interaction sites. METHODS. Peptides were oxidized by using CuSO 4 and H2O2. Aggregation and light-scattering assays of bovine βL-crystallin were conducted at 55°C and 37°C, respectively. Assays were performed in the presence of oxidized and nonoxidized βB3-crystallin peptides and in the presence of α-crystallin. Peptide-induced change in hydrophobicity was determined by bis-ANS (4,4′-dianilino-1,1′ binaphthyl-5,5′ disulfonic acid) binding study. Oxidized βB3-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 βL-crystallin was higher in the presence of oxidized βB3-crystallin peptide than with βL-crystallin, without oxidized peptide and with nonoxidized peptide. Enhanced aggregation was observed despite the presence of α-crystallin in the assay. Furthermore, a significant increase in aggregation and light-scattering was observed in the presence of oxidized βB3-peptide at 37°C. Bis-ANS binding to βL-crystallin treated with oxidized βB3-peptide was two to three times higher than in the controls at 37°C. The oxidized βB3-peptide preferentially interacted with βB2-crystallin. The data were confirmed by mass spectrometric analysis. CONCLUSIONS. Oxidized βB3-peptide interacts with βB2-crystallin and enhances its aggregation and precipitation. Peptide-induced aggregation and increased hydrophobicity of the lens crystallin at 37°C are relevant to crystallin aggregation in the aging lenses.
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U2 - 10.1167/iovs.05-0031
DO - 10.1167/iovs.05-0031
M3 - Article
C2 - 15980243
AN - SCOPUS:23244468386
VL - 46
SP - 2514
EP - 2521
JO - Investigative Ophthalmology and Visual Science
JF - Investigative Ophthalmology and Visual Science
SN - 0146-0404
IS - 7
ER -