Molecular characterization of an insecticide-induced novel glutathione transferase in silkworm

研究成果: ジャーナルへの寄稿記事

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抄録

Background: The glutathione transferase (GST) superfamily is involved in the detoxification of various xenobiotics. We have identified a GST mRNA that was induced in the fat bodies of a silkworm strain exhibiting diazinon resistance and have investigated the enzyme properties of this GST. Methods: A soluble recombinant protein was overexpressed in Escherichia coli. Amino acid residues of interest were changed to alanine by site-directed mutagenesis. Results and conclusions: Phylogenetic analysis of the deduced amino acid sequence indicates that this GST belongs to an unclassified group previously reported in mosquitoes. This enzyme, named bmGSTu, has highly conserved amino acid residues, including Tyr7, Ser12 and Asn50. A recombinant bmGSTu was able to catalyze the biotranslation of glutathione with 1-chloro-2,4-dinitrobenzene, a synthetic substrate of GST. Kinetic analysis of bmGSTu mutants indicated that Tyr7, Ser12 and Asn50 are involved in enzyme function. General significance: These results support the hypothesis that bmGSTu may play a role in insecticide resistance in Bombyx mori.

元の言語英語
ページ(範囲)420-426
ページ数7
ジャーナルBiochimica et Biophysica Acta - General Subjects
1810
発行部数4
DOI
出版物ステータス出版済み - 4 1 2011

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Bombyx
Insecticides
Glutathione Transferase
Amino Acids
Enzymes
Diazinon
Insecticide Resistance
Dinitrochlorobenzene
Detoxification
Mutagenesis
Fat Body
Protein Sequence Analysis
Xenobiotics
Site-Directed Mutagenesis
Culicidae
Recombinant Proteins
Alanine
Escherichia coli
Glutathione
Fats

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology

これを引用

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title = "Molecular characterization of an insecticide-induced novel glutathione transferase in silkworm",
abstract = "Background: The glutathione transferase (GST) superfamily is involved in the detoxification of various xenobiotics. We have identified a GST mRNA that was induced in the fat bodies of a silkworm strain exhibiting diazinon resistance and have investigated the enzyme properties of this GST. Methods: A soluble recombinant protein was overexpressed in Escherichia coli. Amino acid residues of interest were changed to alanine by site-directed mutagenesis. Results and conclusions: Phylogenetic analysis of the deduced amino acid sequence indicates that this GST belongs to an unclassified group previously reported in mosquitoes. This enzyme, named bmGSTu, has highly conserved amino acid residues, including Tyr7, Ser12 and Asn50. A recombinant bmGSTu was able to catalyze the biotranslation of glutathione with 1-chloro-2,4-dinitrobenzene, a synthetic substrate of GST. Kinetic analysis of bmGSTu mutants indicated that Tyr7, Ser12 and Asn50 are involved in enzyme function. General significance: These results support the hypothesis that bmGSTu may play a role in insecticide resistance in Bombyx mori.",
author = "Kohji Yamamoto and Hirofumi Ichinose and Yoichi Aso and Yutaka Banno and Makoto Kimura and Takashi Nakashima",
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T1 - Molecular characterization of an insecticide-induced novel glutathione transferase in silkworm

AU - Yamamoto, Kohji

AU - Ichinose, Hirofumi

AU - Aso, Yoichi

AU - Banno, Yutaka

AU - Kimura, Makoto

AU - Nakashima, Takashi

PY - 2011/4/1

Y1 - 2011/4/1

N2 - Background: The glutathione transferase (GST) superfamily is involved in the detoxification of various xenobiotics. We have identified a GST mRNA that was induced in the fat bodies of a silkworm strain exhibiting diazinon resistance and have investigated the enzyme properties of this GST. Methods: A soluble recombinant protein was overexpressed in Escherichia coli. Amino acid residues of interest were changed to alanine by site-directed mutagenesis. Results and conclusions: Phylogenetic analysis of the deduced amino acid sequence indicates that this GST belongs to an unclassified group previously reported in mosquitoes. This enzyme, named bmGSTu, has highly conserved amino acid residues, including Tyr7, Ser12 and Asn50. A recombinant bmGSTu was able to catalyze the biotranslation of glutathione with 1-chloro-2,4-dinitrobenzene, a synthetic substrate of GST. Kinetic analysis of bmGSTu mutants indicated that Tyr7, Ser12 and Asn50 are involved in enzyme function. General significance: These results support the hypothesis that bmGSTu may play a role in insecticide resistance in Bombyx mori.

AB - Background: The glutathione transferase (GST) superfamily is involved in the detoxification of various xenobiotics. We have identified a GST mRNA that was induced in the fat bodies of a silkworm strain exhibiting diazinon resistance and have investigated the enzyme properties of this GST. Methods: A soluble recombinant protein was overexpressed in Escherichia coli. Amino acid residues of interest were changed to alanine by site-directed mutagenesis. Results and conclusions: Phylogenetic analysis of the deduced amino acid sequence indicates that this GST belongs to an unclassified group previously reported in mosquitoes. This enzyme, named bmGSTu, has highly conserved amino acid residues, including Tyr7, Ser12 and Asn50. A recombinant bmGSTu was able to catalyze the biotranslation of glutathione with 1-chloro-2,4-dinitrobenzene, a synthetic substrate of GST. Kinetic analysis of bmGSTu mutants indicated that Tyr7, Ser12 and Asn50 are involved in enzyme function. General significance: These results support the hypothesis that bmGSTu may play a role in insecticide resistance in Bombyx mori.

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