Sequence of pig lens aldose reductase and electrospray mass spectrometry of non-covalent and covalent complexes.
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).
Jaquinod
M
M
Laboratoire de Spectrométrie de Masse Bio-Organique, URA31 CNRS, Université Louis Pasteur, Strasbourg, France.
Potier
N
N
Klarskov
K
K
Reymann
J M
JM
Sorokine
O
O
Kieffer
S
S
Barth
P
P
Andriantomanga
V
V
Biellmann
J F
JF
Van Dorsselaer
A
A
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
D009249:NADP; D008715:Methionine; D000449:Aldehyde Reductase