title: Resolving isoforms of aldose reductase by preparative isoelectric focusing in the Rotofor.
abstract: We have resolved and characterized isoforms of aldose reductase from bovine and porcine lenses by preparative isoelectric focusing with narrow pH gradients using the Rotofor. Both bovine and porcine lens aldose reductases were resolved as two enzyme isoforms. The bovine isoforms were Mr40400 +/- 445 polypeptides of pI4.71 and 5.19. Porcine isoforms were Mr41500 +/- 450 polypeptides of pI 4.90 and 5.30. Staphylococcus aureus V-8 protease digestion patterns for each set of isoforms were essentially identical and all isoforms probably contain blocked amino terminal amino acids. Antiserum to bovine lens aldose reductase cross-reacted with porcine lens aldose reductase. Each isoform displayed substrate preferences characteristic of mammalian aldose reductases. With purification, both bovine and porcine lens aldose reductases became less sensitive to inhibition by 6-fluoro-spiro-(chroman-4.4'-imidazolidine)-2',5'-dione (sorbinil).
authors.affiliation: Department of Ophthalmology and Visual Sciences, Washington University School of Medicine, St. Louis, MO 63110.
authors.forename: J M
authors.initials: JM
authors.lastname: Petrash
authors.forename: L J
authors.initials: LJ
authors.lastname: DeLucas
authors.forename: E
authors.initials: E
authors.lastname: Bowling
authors.forename: N
authors.initials: N
authors.lastname: Egen
mesh_terms: D000449:Aldehyde Reductase / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D007093:Imidazoles; D048289:Imidazolidines; D007525:Isoelectric Focusing / Q000295:instrumentation*; D007908:Lens, Crystalline / Q000201:enzymology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010449:Peptide Mapping; D012680:Sensitivity and Specificity; D012697:Serine Endopeptidases; D013379:Substrate Specificity; D013552:Swine
chemical_list: D007093:Imidazoles; D048289:Imidazolidines; D000449:Aldehyde Reductase; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; C026411:sorbinil