title: Analysis of UVA-related alterations upon aging of eye lens proteins by mini two-dimensional polyacrylamide gel electrophoresis.
abstract: 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 fractions upon aging. The fractions were obtained from irradiated long-term organ culture lenses and analyzed by mini two-dimensional gel electrophoresis. This micropreparative method followed by computer analysis allows high resolution 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 classes by gel filtration. Moreover, we immunoblotted the gel of the urea-soluble fraction with a specific antibody against the intermediate filament protein vimentin. Upon irradiation of young and adult lenses, alphaA-crystallin and vimentin showed obvious modifications. During aging the susceptibility to irradiation increased when vimentin started to degrade, whereas deamidation of alphaA-crystallin seems to occur.
authors.affiliation: Department of Biochemistry, University of Nijmegen, The Netherlands.
authors.forename: O
authors.initials: O
authors.lastname: Weinreb
authors.forename: F A
authors.initials: FA
authors.lastname: van Rijk
authors.forename: H T
authors.initials: HT
authors.lastname: Steely
authors.forename: A
authors.initials: A
authors.lastname: Dovrat
authors.forename: H
authors.initials: H
authors.lastname: Bloemendal
mesh_terms: D000375:Aging / Q000378:metabolism* / Q000528:radiation effects; D000818:Animals; D002386:Cataract / Q000209:etiology / Q000378:metabolism*; D002417:Cattle; D002850:Chromatography, Gel; D003459:Crystallins / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D007908:Lens, Crystalline / Q000378:metabolism / Q000528:radiation effects*; D009924:Organ Culture Techniques; D011833:Radiation Injuries, Experimental / Q000209:etiology / Q000378:metabolism*; D014466:Ultraviolet Rays / Q000009:adverse effects*
chemical_list: D003459:Crystallins