Yabuki H

References (2)

Title : Crystal structure of the novel haloalkane dehalogenase DatA from Agrobacterium tumefaciens C58 reveals a special halide-stabilizing pair and enantioselectivity mechanism - Guan_2014_Appl.Microbiol.Biotechnol_98_8573
Author(s) : Guan L , Yabuki H , Okai M , Ohtsuka J , Tanokura M
Ref : Applied Microbiology & Biotechnology , 98 :8573 , 2014
Abstract : A novel haloalkane dehalogenase DatA from Agrobacterium tumefaciens C58 belongs to the HLD-II subfamily and hydrolyzes brominated and iodinated compounds, leading to the generation of the corresponding alcohol, a halide ion, and a proton. Because DatA possesses a unique Asn-Tyr pair instead of the Asn-Trp pair conserved among the subfamily members, which was proposed to keep the released halide ion stable, the structural basis for its reaction mechanism should be elucidated. Here, we determined the crystal structures of DatA and its Y109W mutant at 1.70 and 1.95 A, respectively, and confirmed the location of the active site by using its novel competitive inhibitor. The structural information from these two crystal structures and the docking simulation suggested that (i) the replacement of the Asn-Tyr pair with the Asn-Trp pair increases the binding affinity for some halogenated compounds, such as 1,3-dibromopropane, mainly due to the electrostatic interaction between Trp109 and halogenated compounds and the change of substrate-binding mode caused by the interaction and (ii) the primary halide-stabilizing residue is only Asn43 in the wild-type DatA, while Tyr109 is a secondary halide-stabilizing residue. Furthermore, docking simulation using the crystal structures of DatA indicated that its enantioselectivity is determined by the large and small spaces around the halogen-binding site.
ESTHER : Guan_2014_Appl.Microbiol.Biotechnol_98_8573
PubMedSearch : Guan_2014_Appl.Microbiol.Biotechnol_98_8573
PubMedID: 24770384
Gene_locus related to this paper: agrtu-DHAA

Title : Crystallization and preliminary X-ray analysis of the haloalkane dehalogenase DatA from Agrobacterium tumefaciens C58 - Mase_2012_Acta.Crystallogr.Sect.F.Struct.Biol.Cryst.Commun_68_652
Author(s) : Mase T , Yabuki H , Okai M , Ohtsuka J , Imai FL , Nagata Y , Tanokura M
Ref : Acta Crystallographica Sect F Struct Biol Cryst Commun , 68 :652 , 2012
Abstract : Haloalkane dehalogenases are enzymes that catalyze the hydrolytic reaction of a wide variety of haloalkyl substrates to form the corresponding alcohol and hydrogen halide products. DatA from Agrobacterium tumefaciens C58 is a haloalkane dehalogenase that has a unique pair of halide-binding residues, asparagine (Asn43) and tyrosine (Tyr109), instead of the asparagine and tryptophan that are conserved in other members of the subfamily. DatA was expressed in Escherichia coli, purified and crystallized using the sitting-drop vapour-diffusion method with a reservoir solution consisting of 0.1 M CHES pH 8.6, 1.0 M potassium sodium tartrate, 0.2 M lithium sulfate, 0.01 M barium chloride. X-ray diffraction data were collected to 1.70 A resolution. The space group of the crystal was determined as the primitive tetragonal space group P422, with unit-cell parameters a = b = 123.7, c = 88.1 A. The crystal contained two molecules in the asymmetric unit.
ESTHER : Mase_2012_Acta.Crystallogr.Sect.F.Struct.Biol.Cryst.Commun_68_652
PubMedSearch : Mase_2012_Acta.Crystallogr.Sect.F.Struct.Biol.Cryst.Commun_68_652
PubMedID: 22684062