PMID:12356301
| Citation |
Almrud, JJ, Kern, AD, Wang, SC, Czerwinski, RM, Johnson, WH Jr, Murzin, AG, Hackert, ML and Whitman, CP (2002) The crystal structure of YdcE, a 4-oxalocrotonate tautomerase homologue from Escherichia coli, confirms the structural basis for oligomer diversity. Biochemistry 41:12010-24 |
|---|---|
| Abstract |
The tautomerase superfamily consists of three major families represented by 4-oxalocrotonate tautomerase (4-OT), 5-(carboxymethyl)-2-hydroxymuconate isomerase (CHMI), and macrophage migration inhibitory factor (MIF). The members of this superfamily are structurally homologous proteins constructed from a simple beta-alpha-beta fold that share a key mechanistic feature; they use an amino-terminal proline, which has an unusually low pK(a), as the general base in a keto-enol tautomerization. Several new members of the 4-OT family have now been identified using PSI-BLAST and categorized into five subfamilies on the basis of multiple-sequence alignments and the conservation of key catalytic and structural residues. The members of subfamily 5, which includes a hypothetical protein designated YdcE from Escherichia coli, are predicted not to form hexamers. The crystal structure of YdcE has been determined to 1.35 A resolution and confirms that it is a dimer. In addition, YdcE complexed with (E)-2-fluoro-p-hydroxycinnamate, identified as a potent competitive inhibitor of this enzyme, as well as N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid (HEPES) and benzoate are also presented. These latter crystal structures reveal the location of the active site and suggest a mechanism for the observed YdcE-catalyzed tautomerization reaction. The dimeric arrangement of YdcE represents a new structure in the 4-OT family and demonstrates structural diversity within the 4-OT family not previously reported. |
| Links | |
| Keywords |
Amino Acid Sequence; Binding Sites; Cloning, Molecular; Crystallography, X-Ray; Escherichia coli/enzymology; Isomerases/antagonists & inhibitors; Isomerases/chemistry; Models, Molecular; Molecular Sequence Data; Phylogeny; Protein Conformation; Pseudomonas putida/chemistry; Sequence Alignment; Sequence Analysis, Protein |
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