2fus
MUTATIONS OF FUMARASE THAT DISTINGUISH BETWEEN THE ACTIVE SITE AND A NEARBY DICARBOXYLIC ACID BINDING SITE
|
OverviewOverview
Two mutant forms of fumarase C from E. coli have been made using PCR and, recombinant DNA. The recombinant form of the protein included a histidine, arm on the C-terminal facilitating purification. Based on earlier studies, two different carboxylic acid binding sites, labeled A- and B-, were, observed in crystal structures of the wild type and inhibited forms of the, enzyme. A histidine at each of the sites was mutated to an asparagine., H188N at the A-site resulted in a large decrease in specific activity, while the H129N mutation at the B-site had essentially no effect. From the, results, we conclude that the A-site is indeed the active site, and a dual, role for H188 as a potential catalytic base is proposed. Crystal, structures of the two mutant proteins produced some unexpected ... [(full description)]
About this StructureAbout this Structure
2FUS is a [Single protein] structure of sequence from [Escherichia coli] with CIT as [ligand]. Active as [Fumarate hydratase], with EC number [4.2.1.2]. Structure known Active Site: S1. Full crystallographic information is available from [OCA].
ReferenceReference
Mutations of fumarase that distinguish between the active site and a nearby dicarboxylic acid binding site., Weaver T, Lees M, Banaszak L, Protein Sci. 1997 Apr;6(4):834-42. PMID:9098893
Page seeded by OCA on Tue Oct 30 17:14:55 2007