title: Glutamate mutase from Clostridium cochlearium: the structure of a coenzyme B12-dependent enzyme provides new mechanistic insights.
abstract: BACKGROUND
  Glutamate mutase (Glm) equilibrates (S)-glutamate with (2S,3S)-3-methylaspartate. Catalysis proceeds with the homolytic cleavage of the organometallic bond of the cofactor to yield a 5'-desoxyadenosyl radical. This radical then abstracts a hydrogen atom from the protein-bound substrate to initiate the rearrangement reaction. Glm from Clostridium cochlearium is a heterotetrameric molecule consisting of two sigma and two epsilon polypeptide chains.
  
  
  RESULTS
  We have determined the crystal structures of inactive recombinant Glm reconstituted with either cyanocobalamin or methylcobalamin. The molecule shows close similarity to the structure of methylmalonyl CoA mutase (MCM), despite poor sequence similarity between its catalytic epsilon subunit and the corresponding TIM-barrel domain of MCM. Each of the two independent B12 cofactor molecules is associated with a substrate-binding site, which was found to be occupied by a (2S,3S)-tartrate ion. A 1:1 mixture of cofactors with cobalt in oxidation states II and III was observed in both crystal structures of inactive Glm.
  
  
  CONCLUSIONS
  The long axial cobalt-nitrogen bond first observed in the structure of MCM appears to result from a contribution of the species without upper ligand. The tight binding of the tartrate ion conforms to the requirements of tight control of the reactive intermediates and suggests how the enzyme might use the substrate-binding energy to initiate cleavage of the cobalt-carbon bond. The cofactor does not appear to have a participating role during the radical rearrangement reaction.
authors.affiliation: Abteilung für Strukturbiologie, Institut für Physikalische Chemie, Karl-Franzens-Universität, Graz, Austria.
authors.forename: R
authors.initials: R
authors.lastname: Reitzer
authors.forename: K
authors.initials: K
authors.lastname: Gruber
authors.forename: G
authors.initials: G
authors.lastname: Jogl
authors.forename: U G
authors.initials: UG
authors.lastname: Wagner
authors.forename: H
authors.initials: H
authors.lastname: Bothe
authors.forename: W
authors.initials: W
authors.lastname: Buckel
authors.forename: C
authors.initials: C
authors.lastname: Kratky
mesh_terms: D003013:Clostridium / Q000201:enzymology*; D003035:Cobalt / Q000737:chemistry; D003038:Cobamides / Q000378:metabolism*; D019751:Intramolecular Transferases / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D013644:Tartrates / Q000378:metabolism
chemical_list: D003038:Cobamides; D013644:Tartrates; D003035:Cobalt; D019751:Intramolecular Transferases; C021570:methylaspartate mutase; C000913:cobamamide; C029768:tartaric acid