TY - JOUR
T1 - Characterisation of a diazinon-metabolising glutathione S-transferase in the silkworm Bombyx mori by X-ray crystallography and genome editing analysis
AU - Yamamoto, Kohji
AU - Higashiura, Akifumi
AU - Hirowatari, Aiko
AU - Yamada, Naotaka
AU - Tsubota, Takuya
AU - Sezutsu, Hideki
AU - Nakagawa, Atsushi
N1 - Funding Information:
This work was supported by a Grant-in-Aid for Scientific Research (KAKENHI, 17K19272) from the Ministry of Education, Culture, Sports, Science and Technology of Japan and by a research grant for Young Investigators from the Department of Agriculture, Kyushu University. We thank Dr Keiro Uchino (National Agriculture and Food Research Organization, Ibaraki, Japan) for injecting the silkworms, Mr Kaoru Nakamura and Mr Toshihiko Misawa (National Agriculture and Food Research Organization, Ibaraki, Japan) for rearing the silkworms, and Ms Satoko Kawamoto (National Agriculture and Food Research Organization, Ibaraki, Japan) for technical assistance. Crystallographic analysis was performed under the Collaborative Research Program of Institute for Protein Research, Osaka university (CR-17-05). Diffraction data were collected at the Osaka University beamline BL44XU at SPring-8 (Harima, Japan) (Proposal No. 2017A6764 and 2017B6764).
Publisher Copyright:
© 2018, The Author(s).
PY - 2018/12/1
Y1 - 2018/12/1
N2 - Previously, we found an unclassified glutathione S-transferase 2 (bmGSTu2) in the silkworm Bombyx mori that conjugates glutathione to 1-chloro-2,4-dinitrobenzene and also metabolises diazinon, an organophosphate insecticide. Here, we provide a structural and genome-editing characterisation of the diazinon-metabolising glutathione S-transferase in B. mori. The structure of bmGSTu2 was determined at 1.68 Å by X-ray crystallography. Mutation of putative amino acid residues in the substrate-binding site showed that Pro13, Tyr107, Ile118, Phe119, and Phe211 are crucial for enzymatic function. bmGSTu2 gene disruption resulted in a decrease in median lethal dose values to an organophosphate insecticide and a decrease in acetylcholine levels in silkworms. Taken together, these results indicate that bmGSTu2 could metabolise an organophosphate insecticide. Thus, this study provides insights into the physiological role of bmGSTu2 in silkworms, detoxification of organophosphate insecticides, and drug targets for the development of a novel insecticide.
AB - Previously, we found an unclassified glutathione S-transferase 2 (bmGSTu2) in the silkworm Bombyx mori that conjugates glutathione to 1-chloro-2,4-dinitrobenzene and also metabolises diazinon, an organophosphate insecticide. Here, we provide a structural and genome-editing characterisation of the diazinon-metabolising glutathione S-transferase in B. mori. The structure of bmGSTu2 was determined at 1.68 Å by X-ray crystallography. Mutation of putative amino acid residues in the substrate-binding site showed that Pro13, Tyr107, Ile118, Phe119, and Phe211 are crucial for enzymatic function. bmGSTu2 gene disruption resulted in a decrease in median lethal dose values to an organophosphate insecticide and a decrease in acetylcholine levels in silkworms. Taken together, these results indicate that bmGSTu2 could metabolise an organophosphate insecticide. Thus, this study provides insights into the physiological role of bmGSTu2 in silkworms, detoxification of organophosphate insecticides, and drug targets for the development of a novel insecticide.
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U2 - 10.1038/s41598-018-35207-8
DO - 10.1038/s41598-018-35207-8
M3 - Article
C2 - 30443011
AN - SCOPUS:85056662671
SN - 2045-2322
VL - 8
JO - Scientific reports
JF - Scientific reports
IS - 1
M1 - 16835
ER -