Download MendeleyThe structures of Alcaligenes faecalisD-3-hydroxybutyrate dehydrogenase before and after NAD(+) and acetate binding suggest a dynamical reaction mechanism as a member of the SDR family.
Hoque, M.M., Shimizu, S., Hossain, M.T., Yamamoto, T., Imamura, S., Suzuki, K., Tsunoda, M., Amano, H., Sekiguchi, T., Takenaka, A.(2008) Acta Crystallogr D Biol Crystallogr 64: 496-505
- PubMed: 18453685
- DOI: https://doi.org/10.1107/S0907444908004009
- Primary Citation of Related Structures:
2YZ7, 3VDQ - PubMed Abstract:
D-3-Hydroxybutyrate dehydrogenase, which catalyzes the reversible reaction between D-3-hydroxybutyrate and acetoacetate, has been classified into the short-chain dehydrogenase/reductase family and is a useful marker in the assay of diabetes mellitus and/or ketoacidosis. The enzyme from Alcaligenes faecalis was crystallized in the apo form and in the holo form with acetate as a substrate analogue. The crystal structures of both forms were determined at 2.2 angstroms resolution. The enzyme is a tetramer composed of four subunits assembled with noncrystallographic 222 point symmetry. Each subunit has two domains. The principal domain adopts the Rossmann fold essential for nucleotide binding, which is a common feature of the SDR family. NAD+ is bound in a large cleft in the domain. The pyrophosphate group of NAD+ is covered by the small additional domain, which is supported by two extended arms allowing domain movement. In the catalytic site, a water molecule is trapped by the catalytic Tyr155 and Ser142 residues in the vicinity of the bound NAD+ and acetate. The substrate analogue acetate is bound above the nicotinamide plane. A substrate (D-3-hydroxybutylate) bound model can reasonably be constructed by adding two C atoms into the void space between the water O atom and the methyl group of the acetate, suggesting a substrate-bound state before enzymatic reaction occurs. Based on these structural features, a reaction mechanism has been proposed.
Organizational Affiliation:
Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Kanagawa 226-8501, Japan.
