TY - JOUR
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
N1 - Funding Information:
This work was supported in part by a Grant-in-Aid for Scientific Research (KAKENHI; 21780049 ) from the Ministry of Education, Science, Sports and Culture of Japan and by the Mishima Kaiun Memorial Foundation . This work was also supported in part by the National Bioresource Project (silkworm) from the above Ministry. The authors are greatly indebted to Dr. Yuki Nakamura, Dr. Hiroaki Noda and Dr. Kazuei Mita at the National Institute of Agrobiological Sciences for a series of microarray analyses.
PY - 2011/4
Y1 - 2011/4
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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U2 - 10.1016/j.bbagen.2011.01.003
DO - 10.1016/j.bbagen.2011.01.003
M3 - Article
C2 - 21237249
AN - SCOPUS:79551650986
SN - 0304-4165
VL - 1810
SP - 420
EP - 426
JO - Biochimica et Biophysica Acta - General Subjects
JF - Biochimica et Biophysica Acta - General Subjects
IS - 4
ER -