Expression and characterization of a sigma-class glutathione S-transferase of the fall webworm, Hyphantria cunea

Kohji Yamamoto, Hiroshi Fujii, Yoichi Aso, Yutaka Banno, Katsumi Koga

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

A cDNA encoding glutathione S-transferase (GST) of the fall webworm, Hyphantria cunea, was cloned by reverse transcriptase-polymerase chain reaction. The resulting clone (hcGST) was sequenced and deduced for amino acid sequence, which revealed 87, 59, and 42% identities to Sigma-class GSTs from Bombyx mori, Manduca sexta, and Blattella germanica respectively. A recombinant hcGST protein (rhcGST) was functionally overexpressed in Escherichia coli cells in a soluble form and purified to homogeneity. rhcGST retained more than 75% of its original GST activity after incubation at pHs 6 to 11. Incubation for 30 min at temperatures below 50°C scarcely affected the activity. rhcGST was able to catalyze the reaction of glutathione with 1- chloro-2,4-dinitrobenzene, a universal substrate for GST, as well as with 4-hydroxynonenal, a product of lipid peroxidation. We also found that as compared to B. mori Sigma-class GST, rhcGST had a higher affinity for fenitrothion, an organophosphorus insecticide.

Original languageEnglish
Pages (from-to)553-560
Number of pages8
JournalBioscience, Biotechnology and Biochemistry
Volume71
Issue number2
DOIs
Publication statusPublished - Mar 1 2007

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Glutathione Transferase
Recombinant Proteins
Bombyx
Fenitrothion
Manduca
Dinitrochlorobenzene
Polymerase chain reaction
RNA-Directed DNA Polymerase
Insecticides
Reverse Transcriptase Polymerase Chain Reaction
Escherichia coli
Lipid Peroxidation
Glutathione
Amino Acid Sequence
Complementary DNA
Clone Cells
Lipids
Amino Acids
Temperature
Substrates

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

Cite this

Expression and characterization of a sigma-class glutathione S-transferase of the fall webworm, Hyphantria cunea. / Yamamoto, Kohji; Fujii, Hiroshi; Aso, Yoichi; Banno, Yutaka; Koga, Katsumi.

In: Bioscience, Biotechnology and Biochemistry, Vol. 71, No. 2, 01.03.2007, p. 553-560.

Research output: Contribution to journalArticle

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