title: Reactivity of aquacobalamin and reduced cobalamin toward S-nitrosoglutathione and S-nitroso-N-acetylpenicillamine.
abstract: The reactions of aquacobalamin (Cbl(III)H2O, vitamin B12a) and reduced cobalamin (Cbl(II), vitamin B12r) with the nitrosothiols S-nitrosoglutathione (GSNO) and S-nitroso-N-acetylpenicillamine (SNAP) were studied in aqueous solution at pH 7.4. UV-vis and NMR spectroscopic studies and semiquantitative kinetic investigations indicated complex reactivity patterns for the studied reactions. The detailed reaction routes depend on the oxidation state of the cobalt center in cobalamin, as well as on the structure of the nitrosothiol. Reactions of aquacobalamin with GSNO and SNAP involve initial formation of Cbl(III)-RSNO adducts followed by nitrosothiol decomposition via heterolytic S-NO bond cleavage. Formation of Cbl(III)(NO-) as the main cobalamin product indicates that the latter step leads to efficient transfer of the NO- group to the Co(III) center with concomitant oxidation of the nitrosothiol. Considerably faster reactions with Cbl(II) proceed through initial Cbl(II)-RSNO intermediates, which undergo subsequent electron-transfer processes leading to oxidation of the cobalt center and reduction of the nitrosothiol. In the case of GSNO, the overall reaction is fast (k approximately 1.2 x 10(6) M(-1) s(-1)) and leads to formation of glutathionylcobalamin (Cbl(III)SG) and nitrosylcobalamin (Cbl(III)(NO-)) as the final cobalamin products. A mechanism involving the reversible equilibrium Cbl(II) + RSNO <==> Cbl(III)SR + NO is suggested for the reaction on the basis of the obtained kinetic and mechanistic information. The corresponding reaction with SNAP is considerably slower and occurs in two distinct reaction steps, which result in the formation of Cbl(III)(NO-) as the ultimate cobalamin product. The significantly different kinetic and mechanistic features observed for the reaction of GSNO and SNAP illustrate the important influence of the nitrosothiol structure on its reactivity toward metal centers of biomolecules. The potential biological implications of the results are briefly discussed.
authors.affiliation: Faculty of Chemistry, Jagiellonian University, 30060 Krakow, Poland.
authors.forename: Maria
authors.initials: M
authors.lastname: Wolak
authors.forename: Grazyna
authors.initials: G
authors.lastname: Stochel
authors.forename: Rudi
authors.initials: R
authors.lastname: van Eldik
mesh_terms: D015394:Molecular Structure; D026423:S-Nitroso-N-Acetylpenicillamine / Q000737:chemistry*; D026422:S-Nitrosoglutathione / Q000737:chemistry*; D014805:Vitamin B 12 / Q000031:analogs & derivatives* / Q000737:chemistry*
chemical_list: C043960:aquacobalamin; D026422:S-Nitrosoglutathione; D026423:S-Nitroso-N-Acetylpenicillamine; D014805:Vitamin B 12