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.
Subject(s)
Animals , Amino Acid Sequence , 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/chemistryABSTRACT
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.
Subject(s)
Animals , Amino Acid Sequence , 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/chemistryABSTRACT
Objective To assess metabolic state in patients with Roux-y sigmoid neobladder.Methods The study comprised 33 patients(21 men and 12 women) with Roux-y sigmoid neobladder after oneological sur-gery.All enrolled patients were treated by the same protocol.Before and after withdrawing the catheter , serum e-lectrolytes, ereatinine and urea were analysed and used to assess the effect.Results All 33 patients were evalu-able.Before and after withdrawing the catheter , serum electrolytes, creatinine and urea were normal values and there was no signifcant difference(P >0.05).Three patients developed mild metabolic acidosis.Conclusions The Roux-y sigmoid neobladder is a feasible , safe and effective method for continent urinary diversion.This surgi-cal technique had no signifcant effect on metabolic state.