Carboxylate-substituted radicals from phenylselenide derivatives. Designs on models for coenzyme B12-dependent enzyme-catalyzed rearrangements

Laser flash photolysis (266 nm) of alpha- and beta-phenylselenyl esters, carboxylic acids, and carboxylates in aqueous acetonitrile media gave the corresponding radicals by homolytic cleavage of the phenylselenyl groups. In the beta-substituted systems, acid and carboxylate radicals reacted in intramolecular reporter reactions with approximately equal rate constants. For the alpha-substituted systems, an ester- and carboxylic acid-substituted radical reacted in an intramolecular reporter reaction with the same rate constants, but the analogous alpha-carboxylate radical, a radical anion, reacted an order of magnitude less rapidly and with an activation energy that is 3 kcal/mol greater than that found for analogues. A kinetic titration of the equilibrating alpha-acid and alpha-carboxylate radicals gave pKa = 4.6. The results indicate that alpha-ester and alpha-carboxylic acid radicals are unlikely to be appropriate models for alpha-carboxylate radicals, the intermediates formed in a large subset of coenzyme B12-dependent enzyme-catalyzed reactions.     

New developments in the field of vitamin B12: enzymatic reactions dependent upon corrins and coenzyme B12.

Simple corrins such as vitamin B₁₂ and vitamin B₁₂ coenzyme catalyze a variety of unusual enzymatic reactions of which some are still without analogy in organic or organometallic chemistry. The mechanisms of these reactions are currently the subject of lively discussion. The present review focuses attention on new ideas about the mode of action of vitamin B₁₂ coenzymes in enzymatic reactions.     

The Co(I) induced methylmalonyl-succinyl rearrangement in a model for the coenzyme B12 dependent methylmalonyl-CoA mutase

The rearrangement of 2-bromomethyl-2-methylmonothiomalonates to succinyl derivatives was found to take place in quantitative yields in the presence of one molar equivalent of Co(I) generated by the reduction of heptamethyl Co(II)yrinate perchlorate with NaBH4 or electrochemically. The chiral thiomalonate gave racemic succinate.     

Ligand substitution behavior of a simple model for coenzyme B12

The ligand substitution reactions of trans-[CoIII(en)2(Me)H2O]2+, a simple model for coenzyme B12, were studied for cyanide and imidazole as entering nucleophiles. It was found that these nucleophiles displace the coordinated water molecule trans to the methyl group and form the six-coordinate complex trans-[Co(en)2(Me)L]. The complex-formation constants for cyanide and imidazole were found to be (8.3 +/- 0.7) x 10(4) and 24.5 +/- 2.2 M-1 at 10 and 12 degrees C, respectively. The second-order rate constants for the substitution of water were found to be (3.3 +/- 0.1) x 10(3) and 198 +/- 13 M-1 s-1 at 25 degrees C for cyanide and imidazole, respectively. From temperature and pressure dependence studies, the activation parameters delta H++, delta S++, and delta V++ for the reaction of trans-[CoIII(en)2(Me)H2O]2+ with cyanide were found to be 50 +/- 4 kJ mol-1, 0 +/- 16 J K-1 mol-1, and +7.0 +/- 0.6 cm3 mol-1, respectively, compared to 53 +/- 2 kJ mol-1, -22 +/- 7 J K-1 mol-1, and +4.7 +/- 0.1 cm3 mol-1 for the reaction with imidazole. On the basis of reported activation volumes, these reactions follow a dissociative mechanism in which the entering nucleophile could be weakly bound in the transition state.     

How the Co-C bond is cleaved in coenzyme B12 enzymes: a theoretical study

The homolytic cleavage of the organometallic Co-C bond in vitamin B12-dependent enzymes is accelerated by a factor of approximately 10(12) in the protein compared to that of the isolated cofactor in aqueous solution. To understand this much debated effect, we have studied the Co-C bond cleavage in the enzyme glutamate mutase with combined quantum and molecular mechanics methods. We show that the calculated bond dissociation energy (BDE) of the Co-C bond in adenosyl cobalamin is reduced by 135 kJ/mol in the enzyme. This catalytic effect can be divided into four terms. First, the adenosine radical is kept within 4.2 angstroms of the Co ion in the enzyme, which decreases the BDE by 20 kJ/mol. Second, the surrounding enzyme stabilizes the dissociated state by 42 kJ/mol using electrostatic and van der Waals interactions. Third, the protein itself is stabilized by 11 kJ/mol in the dissociated state. Finally, the coenzyme is geometrically distorted by the protein, and this distortion is 61 kJ/mol larger in the Co(III) state. This deformation of the coenzyme is caused mainly by steric interactions, and it is especially the ribose moiety and the Co-C5'-C4' angle that are distorted. Without the polar ribose group, the catalytic effect is much smaller, e.g. only 42 kJ/mol for methyl cobalamin. The deformation of the coenzyme is caused mainly by the substrate, a side chain of the coenzyme itself, and a few residues around the adenosine part of the coenzyme.     

Stabilisation of methylene radicals by cob(II)alamin in coenzyme B12 dependent mutases

Coenzyme B12 initiates radical chemistry in two types of enzymatic reactions, the irreversible eliminases (e.g., diol dehydratases) and the reversible mutases (e.g., methylmalonyl-CoA mutase). Whereas eliminases that use radical generators other than coenzyme B12 are known, no alternative coenzyme B12 independent mutases have been detected for substrates in which a methyl group is reversibly converted to a methylene radical. We predict that such mutases do not exist. However, coenzyme B12 independent pathways have been detected that circumvent the need for glutamate, beta-lysine or methylmalonyl-CoA mutases by proceeding via different intermediates. In humans the methylcitrate cycle, which is ostensibly an alternative to the coenzyme B12 dependent methylmalonyl-CoA pathway for propionate oxidation, is not used because it would interfere with the Krebs cycle and thereby compromise the high-energy requirement of the nervous system. In the diol dehydratases the 5'-deoxyadenosyl radical generated by homolysis of the carbon-cobalt bond of coenzyme B12 moves about 10 A away from the cobalt atom in cob(II)alamin. The substrate and product radicals are generated at a similar distance from cob(II)alamin, which acts solely as spectator of the catalysis. In glutamate and methylmalonyl-CoA mutases the 5'-deoxyadenosyl radical remains within 3-4 A of the cobalt atom, with the substrate and product radicals approximately 3 A further away. It is suggested that cob(II)alamin acts as a conductor by stabilising both the 5'-deoxyadenosyl radical and the product-related methylene radicals.     

A nonenzymic model for the coenzyme B12-dependent isomerization of methylmalonyl-SCoA to suiccinyl-SCoA

Dimethyl bromomethylmalonate (IV) reacts with vitamin B_12s in aqueous solution yielding a relatively unstable carbon-cobalt bonded adduct V, which shows visible spectra in good accord with expectation. The adduct V was allowed to decompose in water, in the dark, at room temperature and at physiological pH. Three products: succinic acid (VI), methylmalonic acid (VIII) and malonic acid (VII) were formed in 3, 18, and 13% yields respectively. Isolation of the succinic acid rearrangement product provides support for the intermediacy of the carbon-cobalt bond in the coenzyme B₁₂ dependent enzymic carbon-skeleton rearrangement of methylmalonyl-SCoA to succinyl-SCoA.     

A chromophore-supported structural and functional model of dinuclear copper enzymes,for facilitating mechanism of action studies

Type III dicopper centres are the heart of the reactive sites of enzymes that catalyze the oxidation of catechols. Numerous synthetic model complexes have been prepared to uncover the fundamental chemistry involved in these processes, but progress is still lagging much behind that for heme enzymes. One reason is that the latter gain very much from the informative spectroscopic features of their porphyrin-based metal-chelating ligand. We now introduce sapphyrin-chelated dicopper complexes and show that they may be isolated in different oxidation states and coordination geometries, with distinctive colors and electronic spectra due to the heme-like ligands. The dicopper(i) complex 1-Cu2 was characterized by ¹H and ¹⁹F NMR spectroscopy of the metal-chelating sapphyrin, the oxygenated dicopper(ii) complex 1-Cu2O2 by EPR, and crystallographic data was obtained for the tetracopper(ii)-bis-sapphyrin complex [1-Cu2O2]2. This uncovered a non-heme [Cu₄(OH)₄]⁴⁻ cluster, held together with the aid of two sapphyrin ligands, with structural features reminiscent of those of catechol oxidase. Biomimetic activity was demonstrated by the 1-Cu2O2 catalyzed aerobic oxidation of catechol to quinone; the sapphyrin ligand aided very much in gaining information about reactive intermediates and the rate-limiting step of the reaction.

Di-copper chelation by sapphyrin facilitates reaction mechanism investigations and characterization of reactive intermediates regarding biomimetic catechol oxidation.     

Radiative lifetimes for the B1sigma(u)+ and C1pi states of the H2 molecule

With RKR electronic potentials for the B1sigma(u)+, C1pi(u) and X1sigma(u)+ states in conjunction with Huffaker's correction and appropriate asymptotic functions, the unperturbed radiative lifetimes of rovibrational levels of the B1sigma(u)+ and C1pi(u) states of H2 are calculated. Comparison with previous calculations is presented. Better lifetimes for B1sigma(u)+ are obtained in present work.     

P-O donor action from carboxylate anions with phosphorus in the presence of hydrogen bonding. A model for phosphoryl-transfer enzymes

A series of phosphorus compounds (1-3) containing anionic carboxylate groups were synthesized by treatment of the respective neutral precursor acid forms B-D with amines, which also served to introduce hydrogen-bonding interactions. The compounds, subjected to X-ray structure analysis, resulted in hexacoordinated anionic phosphoranates 1A and 1B, a pseudo-trigonal-bipyramidal anionic phosphine (2), and a trigonal-bipyramidal anionic phosphine oxide (3). The structures revealed that P-O donor coordination was present in all members of the anionic series 1-3 and resulted in stronger interactions than existed in the precursor neutral acid forms B-D as measured by the presence of shorter P-O distances. Evaluation of the energies of the donor interactions relative to the energies of the hydrogen bonds that were present showed that the donor energies now exceeded the hydrogen bond strengths. (31)P chemical shifts indicated that the basic coordination geometries were retained in solution. Both 1A and 1B are chiral and exist as racemates. The results suggest that mechanisms of phosphoryl-transfer enzymes should benefit by taking into account donor interactions at phosphorus by residues at active sites in addition to the inclusion of hydrogen bonding. Reference is made to specific phosphoryl-transfer enzymes.