title: Light scattering and photocrosslinking in the calf lens crystallins gamma-II, III and IV.
abstract: The calf lens proteins gamma-II, -III and -IV crystallin have been photolyzed in pH 7.5 phosphate buffer solution at 25 degrees C. The photolysis light source was either a xenon arc lamp/monochromator system set to pass 290 +/- 5 nm or a nitrogen laser operating at 337.1 nm. Photolysis experiments at 337.1 nm were done both in the presence and absence of added 1.0 x 10(-4) M N-formylkynurenine (NFK). In addition, 1 x 10(-5) M riboflavin was added as a photosensitizer in a few of the experiments. All solutions were 1.0 mg ml-1 protein, and 1.0 ml of solution was irradiated for periods ranging from 10 min to 3 hr. During the 337.1 nm irradiations, the turbidity of the protein solutions was continuously monitored using a He-Ne laser at 632.8 nm. Progress of the 290 nm irradiations was monitored by observing the loss of tryptophan fluorescence for each of the gamma crystallin proteins. The rate of growth of light scattering, upon 337.1 nm irradiation, was greatest for gamma-IV. Addition of NFK caused the rates of growth of UV-induced light scattering of all three gamma crystallins to increase significantly. These rates were in the order: gamma-IV much greater than gamma-III greater than gamma-II. Following UV exposure, the protein solutions were analyzed using UV-visible absorption spectroscopy and SDS-PAGE. Irradiated gamma crystallin solutions showed increased optical density throughout the visible region, resulting from solution turbidity.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: School of Chemistry, Georgia Institute of Technology, Atlanta 30332.
authors.forename: M L
authors.initials: ML
authors.lastname: Walker
authors.forename: R F
authors.initials: RF
authors.lastname: Borkman
mesh_terms: D000818:Animals; D002417:Cattle; D003459:Crystallins / Q000378:metabolism / Q000528:radiation effects*; D008027:Light; D010782:Photolysis; D012542:Scattering, Radiation; D014466:Ultraviolet Rays
chemical_list: D003459:Crystallins