PMID:14718529
| Citation |
Forouhar, F, Lee, I, Benach, J, Kulkarni, K, Xiao, R, Acton, TB, Montelione, GT and Tong, L (2004) A novel NAD-binding protein revealed by the crystal structure of 2,3-diketo-L-gulonate reductase (YiaK). J. Biol. Chem. 279:13148-55 |
|---|---|
| Abstract |
Escherichia coli YiaK catalyzes the reduction of 2,3-diketo-L-gulonate in the presence of NADH. It belongs to a large family of oxidoreductases that is conserved in archaea, bacteria, and eukaryotes but shows no sequence homology to other proteins. We report here the crystal structures at up to 2.0-A resolution of YiaK alone and in complex with NAD-tartrate. YiaK has a new polypeptide backbone fold and a novel mode of recognizing the NAD cofactor. In addition, NAD is bound in an unusual conformation, at the interface of a dimer of the enzyme. The crystallographic analysis unexpectedly revealed the binding of tartrate in the active site. Enzyme kinetics studies confirm that tartrate and the related D-malate are inhibitors of YiaK. In contrast to most other enzymes where substrate binding produces a more closed conformation, the binding of NAD-tartrate to YiaK produces a more open active site. The free enzyme conformation is incompatible with NAD binding. His(44) is likely the catalytic residue of the enzyme. |
| Links |
PubMed Online version:10.1074/jbc.M313580200 |
| Keywords |
Amino Acid Sequence; Binding Sites; Catalytic Domain; Crystallography, X-Ray; Dimerization; Electrons; Escherichia coli/metabolism; Escherichia coli Proteins/chemistry; Escherichia coli Proteins/metabolism; Kinetics; Models, Chemical; Models, Molecular; Molecular Sequence Data; NAD/chemistry; Protein Binding; Protein Conformation; Protein Structure, Tertiary; Sequence Homology, Amino Acid; Sugar Acids/chemistry; Sugar Alcohol Dehydrogenases/chemistry; Sugar Alcohol Dehydrogenases/metabolism; Tartrates/chemistry |
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