title: Structural characterization of the catalytic site of a Nilaparvata lugens delta-class glutathione transferase.
abstract: Glutathione transferases (GSTs) are a major class of detoxification enzymes that play a central role in the defense against environmental toxicants and oxidative stress. Here, we studied the crystal structure of a delta-class glutathione transferase from Nilaparvata lugens, nlGSTD, to gain insights into its catalytic mechanism. The structure of nlGSTD in complex with glutathione, determined at a resolution of 1.7Å, revealed that it exists as a dimer and its secondary and tertiary structures are similar to those of other delta-class GSTs. Analysis of a complex between nlGSTD and glutathione showed that the bound glutathione was localized to the glutathione-binding site. Site-directed mutagenesis of nlGSTD mutants indicated that amino acid residues Ser11, His52, Glu66, and Phe119 contribute to catalytic activity.
authors.affiliation: Faculty of Agriculture, Kyushu University Graduate School, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan. Electronic address: yamamok@agr.kyushu-u.ac.jp.
authors.forename: Kohji
authors.initials: K
authors.lastname: Yamamoto
authors.affiliation: Institute for Protein Research, Osaka University, Suita 565-0871, Japan.
authors.forename: Akifumi
authors.initials: A
authors.lastname: Higashiura
authors.affiliation: Faculty of Agriculture, Kyushu University Graduate School, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan.
authors.forename: Md Tofazzal
authors.initials: MT
authors.lastname: Hossain
authors.affiliation: Faculty of Agriculture, Kyushu University Graduate School, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan.
authors.forename: Naotaka
authors.initials: N
authors.lastname: Yamada
authors.affiliation: National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8634, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Shiotsuki
authors.affiliation: Institute for Protein Research, Osaka University, Suita 565-0871, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Nakagawa
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005978:Glutathione / Q000737:chemistry*; D005982:Glutathione Transferase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006430:Hemiptera / Q000737:chemistry* / Q000201:enzymology; D019476:Insect Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D011485:Protein Binding; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013329:Structure-Activity Relationship
chemical_list: D019476:Insect Proteins; D011994:Recombinant Proteins; D005982:Glutathione Transferase; D005978:Glutathione
keywords: Crystal structure; Glutathione; Glutathione transferase; Nilaparvata lugens; Site-directed mutagenesis