Korean J Parasitol.  2015 Feb;53(1):65-75. 10.3347/kjp.2015.53.1.65.

Identification and Molecular Characterization of Parkin in Clonorchis sinensis

• Bai X
• Kim TI
• Lee JY
• Dai F
• Hong SJ

• KMID: 2381798
• DOI: http://doi.org/10.3347/kjp.2015.53.1.65

Abstract

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 Zn2+ 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.

Keyword

Clonorchis sinensis; parkin; ubiquitin ligase; platyhelminth; mtabolism

MeSH Terms

Amino Acid Sequence
Animals
Clonorchis sinensis/*enzymology
Cluster Analysis
Conserved Sequence
DNA, Complementary/genetics
Energy Metabolism
Gene Expression Profiling
Mitochondria/metabolism
Models, Molecular
Molecular Weight
Phylogeny
Protein Conformation
Sequence Homology, Amino Acid
Ubiquitin-Protein Ligases/chemistry/*genetics/*metabolism
DNA, Complementary
Ubiquitin-Protein Ligases