Identification and molecular characterization of Parkin in Clonorchis sinensis.
Clonorchis sinensis habitating in the bile duct of mammals causes clonorchiasis endemic in East Asian countries. Parkin is a RING-between-RING protein and has E3-ubiquitin ligase activity catalyzing ubiquitination and degradation of substrate proteins. A cDNA clone of C. sinensis was predicted to encode a polypeptide homologous to parkin (CsParkin) including 5 domains (Ubl, RING0, RING1, IBR, and RING2). The cysteine and histidine residues binding to Zn(2+) were all conserved and participated in formation of tertiary structural RINGs. Conserved residues were also an E2-binding site in RING1 domain and a catalytic cysteine residue in the RING2 domain. Native CsParkin was determined to have an estimated molecular weight of 45.7 kDa from C. sinensis adults by immunoblotting. CsParkin revealed E3-ubiquitin ligase activity and higher expression in metacercariae than in adults. CsParkin was localized in the locomotive and male reproductive organs of C. sinensis adults, and extensively in metacercariae. Parkin has been found to participate in regulating mitochondrial function and energy metabolism in mammalian cells. From these results, it is suggested that CsParkin play roles in energy metabolism of the locomotive organs, and possibly in protein metabolism of the reproductive organs of C. sinensis.
Department of Medical Environmental Biology, Chung-Ang University College of Medicine, Seoul 156-756, Korea.
Xuelian
X
Bai
Department of Medical Environmental Biology, Chung-Ang University College of Medicine, Seoul 156-756, Korea.
Tae Im
TI
Kim
Department of Medical Environmental Biology, Chung-Ang University College of Medicine, Seoul 156-756, Korea.
Ji-Yun
JY
Lee
Department of Medical Environmental Biology, Chung-Ang University College of Medicine, Seoul 156-756, Korea.
Fuhong
F
Dai
Department of Medical Environmental Biology, Chung-Ang University College of Medicine, Seoul 156-756, Korea.
Sung-Jong
SJ
Hong
D000595:Amino Acid Sequence; D000818:Animals; D003004:Clonorchis sinensis / Q000201:enzymology*; D016000:Cluster Analysis; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000235:genetics; D004734:Energy Metabolism; D020869:Gene Expression Profiling; D008928:Mitochondria / Q000378:metabolism; D008958:Models, Molecular; D008970:Molecular Weight; D010802:Phylogeny; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*
D018076:DNA, Complementary; D044767:Ubiquitin-Protein Ligases
Clonorchis sinensis; mtabolism; parkin; platyhelminth; ubiquitin ligase