TY - JOUR
T1 - Identification of a sigma-class glutathione-S-transferase from the silkworm, Bombyx mori
AU - Yamamoto, K.
AU - Zhang, P. B.
AU - Banno, Y.
AU - Fujii, H.
PY - 2006/12
Y1 - 2006/12
N2 - A cDNA that encodes a glutathione-S-transferase belonging to sigma class (GSTS1) from the silkworm, Bombyx mori was cloned by reverse transcriptase-polymerase chain reaction and sequenced. The deduced amino acid sequence revealed 66%, 48% and 41% identity to sigma-class GSTs from Manduca sexta, Blattella germanica and Anopheles gambiae, respectively. The GSTS1 was also estimated to be close to those GSTs in a phylogenetic tree. GSTS1 mRNA was widely distributed in various tissues. The recombinant GST (rGSTS1) was functionally overexpressed in Escherichia coli in a soluble form, purified to homogeneity and characterized. The pH-optimum of rGSTS1 was around pH 8. The rGSTS1 retained more than 75% of its original activity after incubation at pH 4-9. Incubation for 30 min at temperatures below 40°C did not affect its activity. rGSTS1 was able to catalyse the reaction of glutathione with 1-chloro-2,4-dinitrobenzene, the universal substrate for GST, as well as with 4-hydroxynonenal, the product of lipid peroxidation.
AB - A cDNA that encodes a glutathione-S-transferase belonging to sigma class (GSTS1) from the silkworm, Bombyx mori was cloned by reverse transcriptase-polymerase chain reaction and sequenced. The deduced amino acid sequence revealed 66%, 48% and 41% identity to sigma-class GSTs from Manduca sexta, Blattella germanica and Anopheles gambiae, respectively. The GSTS1 was also estimated to be close to those GSTs in a phylogenetic tree. GSTS1 mRNA was widely distributed in various tissues. The recombinant GST (rGSTS1) was functionally overexpressed in Escherichia coli in a soluble form, purified to homogeneity and characterized. The pH-optimum of rGSTS1 was around pH 8. The rGSTS1 retained more than 75% of its original activity after incubation at pH 4-9. Incubation for 30 min at temperatures below 40°C did not affect its activity. rGSTS1 was able to catalyse the reaction of glutathione with 1-chloro-2,4-dinitrobenzene, the universal substrate for GST, as well as with 4-hydroxynonenal, the product of lipid peroxidation.
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U2 - 10.1111/j.1439-0418.2006.01092.x
DO - 10.1111/j.1439-0418.2006.01092.x
M3 - Article
AN - SCOPUS:33751031609
SN - 0931-2048
VL - 130
SP - 515
EP - 522
JO - Journal of Applied Entomology
JF - Journal of Applied Entomology
IS - 9-10
ER -