O. Weinreb et al., Analysis of UVA-Related alterations upon aging of eye lens proteins by mini two-dimensional polyacrylamide gel electrophoresis, OPHTHAL RES, 32(5), 2000, pp. 195-204
This study is a first approach to identify UVA-related alterations in situ
of bovine eye lens proteins from the water-soluble and urea-soluble fractio
ns upon aging. The fractions were obtained from irradiated long-term organ
culture lenses and analyzed by mini two-dimensional gel electrophoresis. Th
is micropreparative method followed by computer analysis allows high resolu
tion and separation of microgram quantities of proteins and to detect spots
which arose as a consequence of irradiation. To facilitate the analysis we
first separated the water-soluble fraction into the major crystallin class
es by gel filtration. Moreover, we immunoblotted the gel of the urea-solubl
e fraction with a specific antibody against the intermediate filament prote
in vimentin. Upon irradiation of young and adult lenses, alpha A-crystallin
and vimentin showed obvious modifications. During aging the susceptibility
to irradiation increased when vimentin started to degrade, whereas deamida
tion of aA-crystallin seems to occur. Copyright (C) 2000 S.Karger AG.Basel.