title: Sequence of pig lens aldose reductase and electrospray mass spectrometry of non-covalent and covalent complexes.
abstract: The complete sequence of pig lens aldose reductase (EC 1.1.1.21), a member of the nicotinamide coenzyme-dependent aldo-keto reductase super family, was determined by the combined use of data obtained from Edman degradation, fast-atom-bombardment mass spectrometry and electrospray mass spectrometry. The N-terminal residue of human and pig aldose reductase was shown to be acetylated. The assignment of a disulfide bridge (Cys298-Cys303) was obtained by mass spectrometry. Electrospray mass spectrometry has been used for molecular mass measurement of human muscle (35758 +/- 7 Da) and pig lens (35778 +/- 3Da) aldose reductase; using mild ionization conditions, it has also been used to study the reversible interaction involved in a non-covalent complex with NADP+ (36527 +/- 4Da). An alkylating analog of NADP+ (3-chloroacetylpyridine-adenine dinucleotide phosphate) was used as an irreversible inhibitor to investigate the NADP binding site and the mass of the covalent complex was measured (36521 +/- 3 Da).
authors.affiliation: Laboratoire de Spectrométrie de Masse Bio-Organique, URA31 CNRS, Université Louis Pasteur, Strasbourg, France.
authors.forename: M
authors.initials: M
authors.lastname: Jaquinod
authors.forename: N
authors.initials: N
authors.lastname: Potier
authors.forename: K
authors.initials: K
authors.lastname: Klarskov
authors.forename: J M
authors.initials: JM
authors.lastname: Reymann
authors.forename: O
authors.initials: O
authors.lastname: Sorokine
authors.forename: S
authors.initials: S
authors.lastname: Kieffer
authors.forename: P
authors.initials: P
authors.lastname: Barth
authors.forename: V
authors.initials: V
authors.lastname: Andriantomanga
authors.forename: J F
authors.initials: JF
authors.lastname: Biellmann
authors.forename: A
authors.initials: A
authors.lastname: Van Dorsselaer
mesh_terms: D000449:Aldehyde Reductase / Q000737:chemistry* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007908:Lens, Crystalline / Q000201:enzymology*; D013058:Mass Spectrometry; D008715:Methionine / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009132:Muscles / Q000201:enzymology; D009249:NADP / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D016339:Spectrometry, Mass, Fast Atom Bombardment; D013552:Swine
chemical_list: D009249:NADP; D008715:Methionine; D000449:Aldehyde Reductase