Cataract-specific posttranslational modifications and changes in the composition of urea-soluble protein fraction from the rat lens

Lyudmila V. Yanshole, Ivan V. Cherepanov, Olga A. Snytnikova, Vadim V. Yanshole, Renad Z. Sagdeev, Yuri P. Tsentalovich

Результат исследования: Научные публикации в периодических изданияхстатьярецензирование

16 Цитирования (Scopus)

Аннотация

Purpose: To determine age-related changes in the composition of the urea-soluble (US) protein fraction from lenses of senescence-accelerated OXYS (cataract model) and Wistar (control) rats and to establish posttranslational modifications (PTMs) occurring under enhanced oxidative stress in OXYS lenses.

Methods: The identity and the relative abundance of crystallins in the US fractions were determined using two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MS). The identities and the positions of PTMs were established using MS/MS measurements.

Results: Two-dimensional gel electrophoresis maps of US protein fractions were obtained for lenses of 3-, 12-, and 62-week-old Wistar and OXYS rats, and the relative abundance of different isoforms of alpha-, beta-, and gamma-crystallins was determined. beta-Crystallins were the major contributor of the US fraction in 3-week-old lenses (above 50%), gamma-crystallins in 12-week-old lenses (50-60%), and in 62-week-old lenses, the contributions from all three crystallin families leveled out. The major interstrain difference was the elevated level of alpha-crystallins in the US fraction from 12-week-old OXYS lenses. Spots with increased relative abundance in OXYS maps were attributed to the cataract-specific spots of interest. The crystallins from these spots were subjected to MS/MS analysis, and the positions of acetylation, oxidation, deamidation, and phosphorylation were established.

Conclusions: The increased relative abundance of a-crystallins in the US fraction from 12-week-old OXYS lenses points to the fast insolubilization of alpha-crystallins under oxidative stress. Most of the PTMs attributed to the cataract-specific modifications also correspond to alpha-crystallins. These PTMs include oxidation of methionine residues, deamidation of asparagine and glutamine residues, and phosphorylation of serine and threonine residues.

Язык оригиналаанглийский
Страницы (с-по)2196-2208
Число страниц13
ЖурналMolecular vision
Том19
СостояниеОпубликовано - 7 ноя 2013

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