title: Purification and electrospray mass spectrometry of aldose reductase from pig lens.
abstract: Aldose reductase (alditol: NADP+ 1-oxidoreductase, EC 1.1.1.21) has been purified from pig lens to homogeneity by a rapid and efficient three-step procedure involving poly(ethylene glycol) fractionation, ion-exchange chromatography and chromatofocusing. The homogeneity of the purified enzyme was examined by polyacrylamide gel electrophoresis under native and denaturing conditions, by isoelectric focusing and by high-performance liquid chromatography on a size-exclusion column. The highly purified enzyme is a monomeric protein with a molecular mass of 35,775 +/- 3 Da as determined by electrospray mass spectrometry (ESMS). This purification procedure is particularly suited for the preparation of triclinic single crystals of pig lens aldose reductase, which are currently used in X-ray studies of this enzyme.
authors.affiliation: Laboratoire de Chimie Organique Biologique, Université Louis Pasteur, Strasbourg, France.
authors.forename: J M
authors.initials: JM
authors.lastname: Reymann
authors.forename: J M
authors.initials: JM
authors.lastname: Rondeau
authors.forename: P
authors.initials: P
authors.lastname: Barth
authors.forename: M
authors.initials: M
authors.lastname: Jaquinod
authors.forename: A
authors.initials: A
authors.lastname: Van Dorsselaer
authors.forename: J F
authors.initials: JF
authors.lastname: Biellmann
mesh_terms: D000449:Aldehyde Reductase / Q000737:chemistry / Q000302:isolation & purification*; D000818:Animals; D007908:Lens, Crystalline / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods; D013552:Swine
chemical_list: D000449:Aldehyde Reductase