Functional characterization of Clonorchis sinensis choline transporter
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Clonorchis sinensis is commonly found in East Asian countries. Clonorchiasis is prevalent in these countries and can lead to various clinical symptoms. In this study, we used overlap extension polymerase chain reaction (PCR) and the Xenopus laevis oocyte expression system to isolate a cDNA encoding the choline transporter of C. sinensis (CsChT). We subsequently characterized recombinant CsChT. Expression of CsChT in X. laevis oocytes enabled efficient transport of radiolabeled choline, with no detectable uptake of arginine, α-ketoglutarate, p-aminohippurate, taurocholate, and estrone sulfate. Influx and efflux experiments showed that CsChT-mediated choline uptake was time- and sodium-dependent, with no exchange properties. Concentration-dependent analyses of revealed saturable kinetics consistent with the Michaelis–Menten equation, while nonlinear regression analyses revealed a Km value of 8.3 μM and a Vmax of 61.0 pmol/oocyte/h. These findings contribute to widen our understanding of CsChT transport properties and the cascade of choline metabolisms within C. sinensis.
Abstract
Clonorchis sinensis is commonly found in East Asian countries. Clonorchiasis is prevalent in these countries and can lead to various clinical symptoms. In this study, we used overlap extension polymerase chain reaction (PCR) and the Xenopus laevis oocyte expression system to isolate a cDNA encoding the choline transporter of C. sinensis (CsChT). We subsequently characterized recombinant CsChT. Expression of CsChT in X. laevis oocytes enabled efficient transport of radiolabeled choline, with no detectable uptake of arginine, α-ketoglutarate, p-aminohippurate, taurocholate, and estrone sulfate. Influx and efflux experiments showed that CsChT-mediated choline uptake was time- and sodium-dependent, with no exchange properties. Concentration-dependent analyses of revealed saturable kinetics consistent with the Michaelis–Menten equation, while nonlinear regression analyses revealed a Km value of 8.3 μM and a Vmax of 61.0 pmol/oocyte/h. These findings contribute to widen our understanding of CsChT transport properties and the cascade of choline metabolisms within C. sinensis.
참고문헌 (40)
- 김은민, "Infection Status of Freshwater Fish with Metacercariae of Clonorchis sinensis in Korea", Parasites, Hosts and Diseases, 46(4), pp.247~251 (2008), 대한기생충학ㆍ열대의학회
- Qian MB, "The global epidemiology of clonorchiasis and its relation with cholangiocarcinoma", Infect Dis Poverty, 1(1), pp.4~ (2012)[DOI]
- Doanh PN, "Clonorchis sinensis and Opisthorchis spp. in Vietnam : current status and prospects", Trans R Soc Trop Med Hyg, 110(1), pp.13~20 (2016)[DOI]
- Tang ZL, "Current status and perspectives of Clonorchis sinensis and clonorchiasis : epidemiology, pathogenesis, omics, prevention and control", Infect Dis Poverty, 5(1), pp.71~ (2016)[DOI]
- Rim HJ, "Clonorchiasis : an update", J Helminthol, 79(3), pp.269~281 (2005)[DOI]
- Bouvard V, "A review of human carcinogens--part B : biological agents", Lancet Oncol, 10(4), pp.321~332 (2009)[DOI]
- Park SK, "Mechanism of praziquantel action at a parasitic flatworm ion channel", Sci Transl Med, 13(625), pp.eabj5832~ (2021)[DOI]
- Robertson AP, "Ion-channels on parasite muscle : pharmacology and physiology", Invert Neurosci, 7(4), pp.209~217 (2007)[DOI]
- You H, "Acetylcholinesterase and nicotinic acetylcholine receptors in schistosomes and other parasitic helminths", Molecules, 22(9), pp.1550~ (2017)[DOI]
- Hu Y, "The new anthelmintic tribendimidine is an L-type(levamisole and pyrantel)nicotinic acetylcholine receptor agonist", PLoS Negl Trop Dis, 3(8), pp.e499~ (2009)[DOI]
- Robertson AP, "Tribendimidine : mode of action and nAChR subtype selectivity in Ascaris and Oesophagostomum", PLoS Negl Trop Dis, 9(2), pp.e0003495~ (2015)[DOI]
- Ancelin ML, "Choline incorporation by Schistosoma mansoni : distribution of choline metabolites during development and after sexual differentiation", J Parasitol, 73(3), pp.530~535 (1987)
- Guidi A, "Drug effects on metabolic profiles of Schistosoma mansoni adult male parasites detected by 1H-NMR spectroscopy", PLoS Negl Trop Dis, 14(10), pp.e0008767~ (2020)[DOI]
- Niculescu MD, "Dietary choline deficiency alters global and gene-specific DNA methylation in the developing hippocampus of mouse fetal brains", FASEB J, 20(1), pp.43~49 (2006)[DOI]
- Zeisel SH, "Choline : an essential nutrient for public health", Nutr Rev, 67(11), pp.615~623 (2009)[DOI]
- Michel V, "Choline transport for phospholipid synthesis", Exp Biol Med, 231(5), pp.490~504 (2006)[DOI]
- Biagini GA, "Malaria parasite transporters as a drug-delivery strategy", Trends Parasitol, 21(7), pp.299~301 (2005)[DOI]
- Young BW, "Uptake and incorporation of choline by Schistosoma mansoni adults", Mol Biochem Parasitol, 15(2), pp.105~114 (1985)[DOI]
- Macedo JP, "Characterization of choline uptake in Trypanosoma brucei procyclic and bloodstream forms", Mol Biochem Parasitol, 190(1), pp.16~22 (2013)[DOI]
- Giacomini KM, "New and emerging research on solute carrier and ATP binding cassette transporters in drug discovery and development : outlook from the international transporter consortium", Clin Pharmacol Ther, 112(3), pp.540~561 (2022)[DOI]
- Park JH, "Geographical distribution and genetic diversity of Plasmodium vivax reticulocyte binding protein 1a correlates with patient antigenicity", PLoS Negl Trop Dis, 16(6), pp.e0010492~ (2022)[DOI]
- Truant AL, "Comparison of formalin-ethyl ether sedimentation, formalin-ethyl acetate sedimentation, and zinc sulfate flotation techniques for detection of intestinal parasites", J Clin Microbiol, 13(5), pp.882~884 (1981)[DOI]
- Cha SH, "Molecular cloning and characterization of multispecific organic anion transporter 4 expressed in the placenta", J Biol Chem, 275(6), pp.4507~4512 (2000)[DOI]
- Kusuhara H, "Molecular cloning and characterization of a new multispecific organic anion transporter from rat brain", J Biol Chem, 274(19), pp.13675~13680 (1999)[DOI]
- Yamaoka K, "A pharmacokinetic analysis program(multi)for microcomputer", J Pharmacobiodyn, 4(11), pp.879~885 (1981)[DOI]
- Ferguson SM, "Lethal impairment of cholinergic neurotransmission in hemicholinium-3-sensitive choline transporter knockout mice", Proc Natl Acad Sci USA, 101(23), pp.8762~8767 (2004)[DOI]
- Canty DJ, "Lecithin and choline in human health and disease", Nutr Rev, 52(10), pp.327~339 (1994)[DOI]
- Biagini GA, "Characterization of the choline carrier of Plasmodium falciparum : a route for the selective delivery of novel antimalarial drugs", Blood, 104(10), pp.3372~3377 (2004)[DOI]
- Webb RA, "A novel Na+", HCO3--codependent choline transporter in the syncytial epithelium of the cestode Hymenolepis diminuta, 1998, pp.119(2), 553~562
- Saric J, "Metabolic profiling of an Echinostoma caproni infection in the mouse for biomarker discovery", PLoS Negl Trop Dis, 2(7), pp.e254~ (2008)[DOI]
- Saric J, "Systems parasitology : effects of Fasciola hepatica on the neurochemical profile in the rat brain", Mol Syst Biol, 6, pp.396~ (2010)[DOI]
- Haga T, "Molecular properties of the high-affinity choline transporter CHT1", J Biochem, 156(4), pp.181~194 (2014)[DOI]
- Muramatsu I, "Regulation of synaptic acetylcholine concentrations by acetylcholine transport in rat striatal cholinergic transmission", J Neurochem, 143(1), pp.76~86 (2017)[DOI]
- Loffelholz K, "Choline, a precursor of acetylcholine and phospholipids in the brain", Prog Brain Res, 98, pp.197~200 (1993)[DOI]
- 채종일, "Praziquantel Treatment in Trematode and Cestode Infections: An Update", Infection and Chemotherapy, 45(1), pp.32~43 (2013), 대한감염학회[DOI]
- Jung-Min Lee, "Hypersensitive Reaction to Praziquantel in a Clonorchiasis Patient", Parasites, Hosts and Diseases, 49(3), pp.273~275 (2011), 대한기생충학ㆍ열대의학회
- Siqueira LDP, "Schistosomiasis : drugs used and treatment strategies", Acta Trop, 176, pp.179~187 (2017)[DOI]
- Xu LL, "Efficacy and safety of praziquantel, tribendimidine and mebendazole in patients with co-infection of Clonorchis sinensis and other helminths", PLoS Negl Trop Dis, 8(8), pp.e3046~ (2014)[DOI]
- Xiao SH, "Comparative effect of mebendazole, albendazole, tribendimidine, and praziquantel in treatment of rats infected with Clonorchis sinensis", Parasitol Res, 108(3), pp.723~730 (2011)[DOI]
- Okuda T, "Competitive inhibition of the high-affinity choline transporter by tetrahydropyrimidine anthelmintics", Eur J Pharmacol, 898, pp.173986~ (2021)[DOI]
