Molecular cloning and characterization of a paramyosin from Clonorchis sinensis.
Paramyosin is a myofibrillar protein present in helminth parasites and plays multifunctional roles in host-parasite interactions. In this study, we identified the gene encoding paramyosin of Clonorchis sinensis (CsPmy) and characterized biochemical and immunological properties of its recombinant protein. CsPmy showed a high level of sequence identity with paramyosin from other helminth parasites. Recombinant CsPmy (rCsPmy) expressed in bacteria had an approximate molecular weight of 100 kDa and bound both human collagen and complement 9. The protein was constitutively expressed in various developmental stages of the parasite. Imunofluorescence analysis revealed that CsPmy was mainly localized in the tegument, subtegumental muscles, and the muscle layer surrounding the intestine of the parasite. The rCsPmy showed high levels of positive reactions (74.6%, 56/75) against sera from patients with clonorchiasis. Immunization of experimental rats with rCsPmy evoked high levels of IgG production. These results collectively suggest that CsPmy is a multifunctional protein that not only contributes to the muscle layer structure but also to non-muscular functions in host-parasite interactions. Successful induction of host IgG production also suggests that CsPmy can be applied as a diagnostic antigen and/or vaccine candidate for clonorchiasis.
Park
Tae-Joon
TJ
Department of Parasitology, Biomedical Center for Brain Korea 21 and Institute of Health Sciences, Gyeongsang National University School of Medicine, Jinju 660-751, Korea.
Kang
Jung-Mi
JM
Na
Byoung-Kuk
BK
Sohn
Woon-Mok
WM
D000595:Amino Acid Sequence; D000825:Animal Structures / Q000737:chemistry; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D003001:Cloning, Molecular; D003004:Clonorchis sinensis / Q000737:chemistry / Q000235:genetics*; D003094:Collagen / Q000378:metabolism; D003186:Complement C9 / Q000378:metabolism; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D007074:Immunoglobulin G / Q000097:blood; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011485:Protein Binding; D051381:Rats; D017207:Rats, Sprague-Dawley; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014335:Tropomyosin / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
D000909:Antibodies, Helminth; D003186:Complement C9; D015801:Helminth Proteins; D007074:Immunoglobulin G; D014335:Tropomyosin; D003094:Collagen