Jm. Sirovatka et Rg. Finke, COENZYME B-12 CHEMICAL PRECEDENT STUDIES - PROBING THE ROLE OF THE IMIDAZOLE BASE-ON MOTIF FOUND IN B-12-DEPENDENT METHYLMALONYL-COA MUTASE, Journal of the American Chemical Society, 119(13), 1997, pp. 3057-3067
Adenosylcobinamide (AdoCbi(+)) plus N-methylimidazole (N-MeIm), [AdoCb
i . N-MeIm]+BF4-, has been studied with the goal of providing a chemic
al precedent for the benzimidazole base-off, protein histidine base-on
form of adenosylcobalamin (AdoCbl, also coenzyme B-12) found in the r
ecent X-ray crystallographic structural study of methylmalonyl-CoA (MM
CoA) mutase. Specifically, the axial-base binding K-assoc and associat
ed Delta H and Delta S thermodynamic parameters for [AdoCbi . N-MeIm]BF4- have been obtained as well as its Co-C thermolytic cleavage produ
cts and kinetic parameters. The thermodynamic studies reveal that imid
azole is unique among the aromatic nitrogenous bases tested, with a mo
re favorable Delta H = -7.8 +/- 0.4 kcal/mol but a compensatingly less
favorable Delta S = -28 +/- 1 eu when binding to AdoCbi(+). A stronge
r, shorter Co-N (N-MeIm) bond is implied for [AdoCbi . N-MeIm](+) (i.e
., vs pyridine as the axial base). The product studies reveal that imi
dazole changes the mode of Co-C cleavage from greater than or equal to
98% homolysis (for the appended 5,6-dimethylbenzimidazole in AdoCbl)
to similar to 50% homolysis and similar to 50% abiological heterolysis
for [AdoCbi . N-MeIm](+). The kinetic studies demonstrate that both C
o-C homolysis and heterolysis are accelerated by the record amounts of
8- and 350-fold, respectively, vs the reference point of 5,6-dimethyl
benzimidazole base-on AdoCbl (and by a record 870- and 30 700-fold, re
spectively vs the reference point of the solvent ethylene glycol as th
e axial base, [AdoCbi . ethylene glycol](+)). The biological significa
nce of these findings is discussed, notably (i) that the MMCoA mutase
subclass of B-12-dependent enzymes must either (a) utilize or (b) prev
ent Co-C heterolysis and (ii) the expectation that a long, ''ideal'' l
ength Co-N(imidazole) is one key way the enzyme can inhibit Co-C heter
olysis as well as accelerate Co-C homolysis. Also discussed are the st
eric and electronic differences of imidazole vs pyridine axial-ligands
, including the literature of imidazole's pi-bonding interactions. Fin
ally, a brief summary of the needed [AdoCbi . base](+) and [Co(II)Cbi
. base](+) structural, computational, and other additional studies is
presented.