Abstract
Aberrant protein interactions can lead to aggregation and insolubilization, such as occurs during cataract formation. Deamidation, a prevalent age-related modification in the lens of the eye, decreases stability of the major lens proteins, crystallins. The mechanism of deamidation altering interactions between αA-crystallin and βB2-crystallin was investigated by detecting changes in solvent accessibility upon complex formation during heating. Solvent accessibility was determined by measuring hydrogen/deuterium exchange levels of backbone amides by high-resolution mass spectrometry. Deuterium levels in wild type βB2-crystallin increased 50-60% in both domains following complex formation with αA-crystallin. This increased solvent accessibility indicated a general loosening along the backbone amides. Peptides with the greatest deuterium increases were located at the buried monomer-monomer interface, suggesting that the βB2 dimer was disrupted. The only region where the deuterium levels decreased was in βB2 peptide 123-139, containing an outside loop, and may be a potential site of interaction with αA. Mimicking deamidation at the βB2 dimer interface prevented complex formation with αA. When temperatures were lowered, an αA/βB2 Q70E/Q162E complex formed with similar solvent accessibilities as αA/WT βB2. Deamidation did not disrupt specific αA/βB2 interactions but favored aggregation before complex formation with αA. We conclude that deamidation contributes to cataract formation through destabilization of crystallins before they can be rescued by α-crystallin.
Original language | English (US) |
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Pages (from-to) | 48-58 |
Number of pages | 11 |
Journal | Experimental Eye Research |
Volume | 104 |
DOIs | |
State | Published - Nov 2012 |
Keywords
- Age-related modifications
- Alpha-crystallin
- Beta-crystallins
- Cataracts
- Crystallins
- Deamdidation
- Lens
ASJC Scopus subject areas
- Ophthalmology
- Sensory Systems
- Cellular and Molecular Neuroscience