Structural and immunological characteristics of a 28-kilodalton cruzipain-like cysteine protease of Paragonimus westermani expressed in the definitive host stage.
A complete cDNA sequence encoding a 28-kDa cruzipain-like cysteine protease of adult Paragonimus westermani, termed Pw28CCP, was isolated from an adult cDNA library. The cDNA contained a single open reading frame of 975 bp encoding 325 amino acids, which exhibited the structural motif and domain organization characteristic of cysteine proteases of non-cathepsin Bs including a hydrophobic signal sequence, an ERFNIN motif, and essential cysteine residues as well as active sites in the mature catalytic region. Analysis of its phylogenetic position revealed that this novel enzyme belonged to the cruzipain-like cysteine proteases. The sequence of the first 13 amino acids predicted from the mature domain of Pw28CCP was in accord with that determined from the native 28-kDa enzyme purified from the adult worm. Expression of Pw28CCP was observed specifically in juvenile and adult worms, with a location in the intestinal epithelium, suggesting that this enzyme could be secreted and involved in nutrient uptake and immune modulation. The recombinant protein expressed in Escherichia coli was used to assess antigenicity by immunoblotting with sera from patients with active paragonimiasis and from those with other parasitic infections. The resulting sensitivity of 86.2% (56 of 65 samples) and specificity of 98% (147 of 150 samples) suggest its potential as an antigen for use in immunodiagnosis.
Yun
D H
DH
Section of Molecular Parasitology, Department of Molecular Cell Biology, Sungkyunkwan University School of Medicine, Suwon 440-746, Korea.
Chung
J Y
JY
Chung
Y B
YB
Bahk
Y Y
YY
Kang
S Y
SY
Kong
Y
Y
Cho
S Y
SY
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002415:Cats; D003546:Cysteine Endopeptidases / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D017931:DNA Primers / Q000235:genetics; D004285:Dogs; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D017238:Genes, Helminth; D007159:Immunologic Tests; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010237:Paragonimiasis / Q000175:diagnosis / Q000276:immunology; D010238:Paragonimus / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000276:immunology; D010802:Phylogeny; D015800:Protozoan Proteins; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D015800:Protozoan Proteins; D003546:Cysteine Endopeptidases; C063214:cruzipain