| RRC ID |
574
|
| Author |
Yamamoto K, Zhang P, Miake F, Kashige N, Aso Y, Banno Y, Fujii H.
|
| Title |
Cloning, expression and characterization of theta-class glutathione S-transferase from the silkworm, Bombyx mori.
|
| Journal |
Comp Biochem Physiol B Biochem Mol Biol
|
| Abstract |
This study focused on glutathione S-transferase (GST), one of the detoxification enzymes, from the silkworm, Bombyx mori (GSTT1). A cDNA encoding a putative GST was amplified by reverse transcriptase-polymerase chain reaction and sequenced. The deduced amino acid sequence revealed 59%, 57% and 56% identities to theta-class GSTs of Musca domestica, Anopheles gambiae and Drosophila melanogaster, respectively. GSTT1 was also estimated to be close to those GSTs in a phylogenetic tree. Recombinant GST (rGSTT1) was functionally overexpressed in Escherichia coli in a soluble form, purified to homogeneity, and characterized. The pH-optimum of rGSTT1 was broad from pH 4 to 9 and rGSTT1 retained more than 75% of its original activity after incubation at pH 5-11. Incubation for 30 min at temperatures below 50 degrees C also affected the activity insignificantly. The Michaelis constant for 1-chloro-2,4-dinitrobenzene was 0.48 mM.
|
| Volume |
141(3)
|
| Pages |
340-6
|
| Published |
2005-7-1
|
| DOI |
10.1016/j.cbpc.2005.04.012
|
| PII |
S1096-4959(05)00084-9
|
| PMID |
15950511
|
| MeSH |
Amino Acid Sequence
Animals
Bombyx / enzymology*
Cloning, Molecular
DNA, Complementary / genetics
DNA, Complementary / metabolism
Glutathione Transferase / genetics
Glutathione Transferase / metabolism*
Hydrogen-Ion Concentration
Molecular Sequence Data
Recombinant Proteins / genetics
Recombinant Proteins / isolation & purification
Recombinant Proteins / metabolism*
Sequence Homology, Amino Acid
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Temperature
|
| IF |
2.219
|
| Times Cited |
64
|
|
WOS Category
|
ZOOLOGY
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| Resource |
| Silkworms |
p50
EST database |
