Large-scale preparation of gamma-crystallin and fractionation by chromatofocusing.
Large quantities of calf gamma-crystallin can be prepared by a single and rapid salting-out procedure. The final product is indistinguishable from the gamma-crystallin fraction obtained after gel filtration of a 15000 g lens protein supernatant over Sephadex G 200. Further fractionation is achieved by the mild procedure of chromatofocusing yielding six to eight subfractions. The latter have been characterized by polyacrylamide gel electrophoresis (PAGE) in 6 M-urea, SDS-gel electrophoresis, partial digestion with Staphylococcus aureus protease and amino acid analysis.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange / Q000379:methods; D003459:Crystallins / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases; D007525:Isoelectric Focusing / Q000379:methods; D008666:Metalloendopeptidases
D000596:Amino Acids; D003459:Crystallins; D010450:Endopeptidases; D008666:Metalloendopeptidases; C022653:auR protein, Staphylococcus aureus