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Methylation of platinum by methylcobalamin

Conference ·
OSTI ID:6696882

Incubation of micromolar levels of potassium hexachloroplatinate (K/sub 2/PtCl/sub 6/) and methylcobalamin (MeB-12) results in the complete conversion of MeB-12 to aquocobalamin (aquoB-12). Demethylation is optimal at approximately pH 2.0 and is accelerated by the addition of potassium tetrachloroplatinate (K/sub 2/PtCl/sub 4/). The reaction is stoichiometric between MeB-12 and the K/sub 2/PtCl/sub 6/ added (1:1). Isosbestic points at 492, 367, and 335 nm during the course of the reaction indicate that MeB-12 is demethylated to aquoB-12 with no accumulation of corrinoid intermediates. Higher alkylcobalamins and methylcobinamide react at much slower rates compared with MeB-12. Incubation of 40..mu..M K/sub 2/ PtCl/sub 6/ with either 40..mu..M (Me-/sup 14/C)MeB-12 or (Me-/sup 3/H)MeB-12 followed by lyophilization converts 70% of the label to a stable form that is associated with platinum upon subsequent paper chromatography and electrophoresis. There is no preferential loss of /sup 3/H relative to /sup 14/C in the reaction product. Difference spectra indicated that the platinum reaction product had an absorption maximum at 260 nm. When 50 ..mu..moles each of (Me-/sup 14/C)MeB-12 and K/sub 2/PtCl/sub 6/ were reacted and subjected to Sephadex G-15 chromatography, the /sup 14/C label eluted with 260 nm of absorbing material. Further chromatography on Sephadex G-15 and CM-cellulose yielded a labeled ultraviolet-absorbing product with a /sup 14/C/Pt ratio of 1.2. The overall recovery was 36 to 42% on the basis of the /sup 14/C. The /sup 14/C-Pt product has absorption maximums at 260 nm and 208 nm, with a minimum at 240 nm (A/sub 240/ nm/A/sub 260/ nm = 0.5). Proton-nuclear magnetic resonance (NMR) spectroscopy confirmed the presence of an H-C-Pt covalent bonding pattern (J for /sup 1/H, /sup 195/Pt = 78.2 Hz; tau for /sup 194/Pt-Me + /sup 196/Pt-Me = 6.956).

OSTI ID:
6696882
Resource Relation:
Conference: Biological implications of metals in the environment, Richland, WA, Sep 1975
Country of Publication:
United States
Language:
English