Molecular cloning and enzymatic characterization of a class mu glutathione S-transferase of Paragonimus westermani.
Glutathione S-transferase (GST) is a component of a second line of defense against bioreactive radicals derived from host immune attack. Paragonimus westermani causes acute or chronic lung diseases in mammals. A cDNA clone, PwGST#11, of adult P. westermani produced in the present study was 748 bp long and encoded an open reading frame of 217 amino acids with a starting methionine. The molecular mass of this putative polypeptide, Pw26GST, was estimated to be 25.1 kDa with an isoelectric point of 5.7. Pw26GST was homologous with the 26-kDa GSTs of trematodes and vertebrates. Nine of the ten amino acid residues lining the glutathione-binding pocket were conserved. Putative Pw26GST polypeptide was clustered with 26-kDa GSTs of trematodes belonging to the class mu. Recombinant Pw26GST protein generated bacterially, revealed GST enzyme activity toward an universal and class mu-specific substrates. Mouse antisera to recombinant Pw26GST protein recognized native 26-kDa GST of P. westermani but not the GSTs of any other trematodes. Collectively, Pw26GST was found to be a member of class mu GSTs of P. westermani.
Kim
Tae Yun
TY
Department of Infection Biology, College of Medicine, Chungnam National University, Daejeon, 301-747, South Korea.
Lee
Ji-Yun
JY
Kim
Tae Im
TI
Moon
Ki Ho
KH
Kang
Shin-Yong
SY
Hong
Sung-Jong
SJ
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D003001:Cloning, Molecular; D016000:Cluster Analysis; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D018509:DNA, Helminth / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D005982:Glutathione Transferase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D048492:Paragonimus westermani / Q000201:enzymology* / Q000235:genetics; D010802:Phylogeny; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
D018076:DNA, Complementary; D018509:DNA, Helminth; D011994:Recombinant Proteins; D005982:Glutathione Transferase