Molecular cloning and immunological characterization of phosphoglycerate kinase from Clonorchis sinensis.
The parasite Clonorchis sinensis was determined to utilize a large amount of external glucose to carry its energy metabolism. Phosphoglycerate kinase (PGK), a glycolytic enzyme, found in many parasites, has been identified as one of the candidate molecules distinguished from human counterparts for vaccine and drug developments. A cDNA clone purified by screening a C. sinensis cDNA library using a heterologous cDNA probe encoded a putative peptide of 415 amino acids with over 60% identities with PGKs from a number of animals. The putative peptides revealed domains corresponding to 12 beta-sheets and inner loops forming a substrate-binding cleft of animal PGKs. The gene product was overexpressed in Escherichia coli and showed a PGK-like enzyme activity. A polyclonal antibody raised against the recombinant C. sinensis PGK was specific to native C. sinensis PGK and localized it to the muscular tissue and tegument of the adult flukes. The C. sinensis PGK elicited antibodies in C. sinensis-infected rabbits. Therefore, it is proposed that C. sinensis PGK could be used as an immunoreagent in the serodiagnosis for clonorchiasis.
Hong
S J
SJ
Department of Parasitology and Pathology, Chung-Ang University College of Medicine, Tongjak-gu, 156-756, Seoul, South Korea. hongsj@cau.ac.kr
Seong
K Y
KY
Sohn
W M
WM
Song
K Y
KY
D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D003001:Cloning, Molecular; D003003:Clonorchiasis / Q000175:diagnosis / Q000469:parasitology; D003004:Clonorchis sinensis / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000276:immunology; D018076:DNA, Complementary / Q000235:genetics; D004285:Dogs; D007150:Immunohistochemistry; D051379:Mice; D008969:Molecular Sequence Data; D010735:Phosphoglycerate Kinase / Q000737:chemistry / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA
D000909:Antibodies, Helminth; D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D010735:Phosphoglycerate Kinase