Fumarate reductase: structural and mechanistic insights from the catalytic reduction of 2-methylfumarate.
The soluble fumarate reductase (FR) from Shewanella frigidimarina can catalyse the reduction of 2-methylfumarate with a k(cat) of 9.0 s(-1) and a K(M) of 32 microM. This produces the chiral molecule 2-methylsuccinate. Here, we present the structure of FR to a resolution of 1.5 A with 2-methylfumarate bound at the active site. The mode of binding of 2-methylfumarate allows us to predict the stereochemistry of the product as (S)-2-methylsuccinate. To test this prediction we have analysed the product stereochemistry by circular dichroism spectroscopy and confirmed the production of (S)-2-methylsuccinate.
EaStCHEM, School of Chemistry, University of Edinburgh, West Mains Road, Edinburgh EH9 3JJ, UK.
Caroline
C
Wardrope
Christopher G
CG
Mowat
Malcolm D
MD
Walkinshaw
Graeme A
GA
Reid
Stephen K
SK
Chapman
D002384:Catalysis; D002942:Circular Dichroism; D003461:Crystallography; D005650:Fumarates / Q000737:chemistry*; D008298:Maleates / Q000737:chemistry*; D010084:Oxidation-Reduction; D011487:Protein Conformation; D020592:Shewanella / Q000201:enzymology*; D013385:Succinate Dehydrogenase / Q000737:chemistry*; D013386:Succinates / Q000737:chemistry
D005650:Fumarates; D008298:Maleates; D013386:Succinates; C073341:citraconic acid; D013385:Succinate Dehydrogenase; C041105:monomethyl succinate