title: Spectroscopic and computational studies of cobalamin species with variable lower axial ligation: implications for the mechanism of Co-C bond activation by class I cobalamin-dependent isomerases.
abstract: 5'-deoxyadenosylcobalamin (coenzyme B12, AdoCbl) serves as the cofactor for several enzymes that play important roles in fermentation and catabolism. All of these enzymes initiate catalysis by promoting homolytic cleavage of the cofactor's Co-C bond in response to substrate binding to their active sites. Despite considerable research efforts, the role of the lower axial ligand in facilitating Co-C bond homolysis remains incompletely understood. In the present study, we characterized several derivatives of AdoCbl and its one-electron reduced form, Co(II)Cbl, by using electronic absorption and magnetic circular dichroism spectroscopies. To complement our experimental data, we performed computations on these species, as well as additional Co(II)Cbl analogues. The geometries of all species investigated were optimized using a quantum mechanics/molecular mechanics method, and the optimized geometries were used to compute absorption spectra with time-dependent density functional theory. Collectively, our results indicate that a reduction in the basicity of the lower axial ligand causes changes to the cofactor's electronic structure in the Co(II) state that replicate the effects seen upon binding of Co(II)Cbl to Class I isomerases, which replace the lower axial dimethylbenzimidazole ligand of AdoCbl with a protein-derived histidine (His) residue. Such a reduction of the basicity of the His ligand in the enzyme active site may be achieved through proton uptake by the catalytic triad of conserved residues, DXHXGXK, during Co-C bond homolysis.
authors.affiliation: †Department of Chemistry, University of Wisconsin-Madison, Madison, Wisconsin 53706, United States.
authors.forename: Karen S
authors.initials: KS
authors.lastname: Conrad
authors.affiliation: †Department of Chemistry, University of Wisconsin-Madison, Madison, Wisconsin 53706, United States.
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Jordan
authors.affiliation: §Department of Chemistry and Biochemistry, Ohio University, Athens, Ohio 45701, United States.
authors.forename: Kenneth L
authors.initials: KL
authors.lastname: Brown
authors.affiliation: †Department of Chemistry, University of Wisconsin-Madison, Madison, Wisconsin 53706, United States.
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Brunold
mesh_terms: D002244:Carbon / Q000737:chemistry* / Q000378:metabolism; D002942:Circular Dichroism; D003035:Cobalt / Q000737:chemistry* / Q000378:metabolism; D003038:Cobamides / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D015394:Molecular Structure; D009942:Organometallic Compounds / Q000737:chemistry* / Q000378:metabolism; D011789:Quantum Theory
chemical_list: D003038:Cobamides; D009942:Organometallic Compounds; D003035:Cobalt; D002244:Carbon; C000913:cobamamide