Functional characterization of glucose transporter 4 involved in glucose uptake in Clonorchis sinensis
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Clonorchis sinensis, which causes clonorchiasis, is prevalent in East Asian countries and poses notable health risks, including bile duct complications. Although praziquantel is the primary treatment for the disease, the emerging resistance among trematodes highlights the need for alternative strategies. Understanding the nutrient uptake mechanisms in trematodes, including C. sinensis, is crucial for developing future effective treatments. This study aimed to characterize the function of C. sinensis glucose transporter 4 (CsGTP4) and determine its role in nutrient uptake employing synthesized cDNA of adult C. sinensis worms. The functional characterization of CsGTP4 involved injecting its cRNA into Xenopus laevis oocytes and analyzing the deoxy-D-glucose uptake levels. The results demonstrated that deoxy-D-glucose uptake depended on the deoxy-D-glucose incubation and CsGTP4 expression time, but not sodium-dependent. The concentration-dependent uptake followed the Michaelis–Menten equation, with a Km value of 2.7 mM and a Vmax value of 476 pmol/oocyte/h based on the Lineweaver–Burk analysis. No uptake of radiolabeled α-ketoglutarate, p-aminohippurate, taurocholate, arginine, or carnitine was observed. The uptake of deoxy-D-glucose by CsGTP4 was significantly inhibited by unlabeled glucose and galactose in a concentration-dependent manner. It was significantly inhibited under strongly acidic and basic conditions. These insights into the glucose uptake kinetics and pH dependency of CsGTP4 provide a deeper understanding of nutrient acquisition in trematodes. This study contributes to the development of novel antiparasitic agents, addressing a considerable socioeconomic challenge in affected regions.
Abstract
Clonorchis sinensis, which causes clonorchiasis, is prevalent in East Asian countries and poses notable health risks, including bile duct complications. Although praziquantel is the primary treatment for the disease, the emerging resistance among trematodes highlights the need for alternative strategies. Understanding the nutrient uptake mechanisms in trematodes, including C. sinensis, is crucial for developing future effective treatments. This study aimed to characterize the function of C. sinensis glucose transporter 4 (CsGTP4) and determine its role in nutrient uptake employing synthesized cDNA of adult C. sinensis worms. The functional characterization of CsGTP4 involved injecting its cRNA into Xenopus laevis oocytes and analyzing the deoxy-D-glucose uptake levels. The results demonstrated that deoxy-D-glucose uptake depended on the deoxy-D-glucose incubation and CsGTP4 expression time, but not sodium-dependent. The concentration-dependent uptake followed the Michaelis–Menten equation, with a Km value of 2.7 mM and a Vmax value of 476 pmol/oocyte/h based on the Lineweaver–Burk analysis. No uptake of radiolabeled α-ketoglutarate, p-aminohippurate, taurocholate, arginine, or carnitine was observed. The uptake of deoxy-D-glucose by CsGTP4 was significantly inhibited by unlabeled glucose and galactose in a concentration-dependent manner. It was significantly inhibited under strongly acidic and basic conditions. These insights into the glucose uptake kinetics and pH dependency of CsGTP4 provide a deeper understanding of nutrient acquisition in trematodes. This study contributes to the development of novel antiparasitic agents, addressing a considerable socioeconomic challenge in affected regions.
참고문헌 (37)
- Kim EM, "Infection status of freshwater fish with metacercariae of Clonorchis sinensis in Korea", Korean J Parasitol, 46(4), pp.247~251 (2008)[DOI]
- 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.321~322 (2009)[DOI]
- Park SK, "Mechanism of praziquantel action at a parasitic flatworm ion channel", Sci Transl Med, 13(625), pp.eabj5832~ (2021)
- Scheepers A, "The glucose transporter families SGLT and GLUT : molecular basis of normal and aberrant function", JPEN J Parenter Enteral Nutr, 28(5), pp.364~731 (2004)[DOI]
- Zhao FQ, "Functional properties and genomics of glucose transporters", Curr Genomics, 8(2), pp.113~128 (2007)[DOI]
- Bell GI, "Molecular biology of mammalian glucose transporters", Diabetes Care, 13(3), pp.198~208 (1990)[DOI]
- Carruthers A, "Facilitated diffusion of glucose", Physiol Rev, 70(4), pp.1135~1176 (1990)[DOI]
- Turk E, "Membrane topology motifs in the SGLT cotransporter family", J Membr Biol, 159(1), pp.1~20 (1997)[DOI]
- Joost HG, "The extended GLUT-family of sugar", polyol transport facilitators : nomenclature, sequence characteristics, and potential function of its novel members(review), 2001, pp.18(4), 247~256
- Skelly PJ, "Cloning, characterization, and functional expression of cDNAs encoding glucose transporter proteins from the human parasite Schistosoma mansoni", J Biol Chem, 269(6), pp.4247~4253 (1994)
- Dean S, "Basic biology of trypanosoma brucei with reference to the development of chemotherapies", Curr Pharm Des, 27(14), pp.1650~1670 (2021)[DOI]
- Won JY, "Functional characterization of Plasmodium vivax hexose transporter 1", Front Cell Infect Microbiol, 13, pp.1321240~ (2023)[DOI]
- Wright EM, "The sodium", glucose cotransport family SLC5, 2004, pp.447(5), 510~518
- Asch HL, "Membrane transport in Schistosoma mansoni : transport of amino acids by adult males", Exp Parasitol, 38(1), pp.123~135 (1975)[DOI]
- Asch HL, "Transtegumental absorption of amino acids by male Schistosoma mansoni", J Parasitol, 61(2), pp.378~379 (1975)
- Ahn SK, "Molecular cloning and functional characterization of a glucose transporter(CsGLUT)in Clonorchis sinensis", Parasitol Res, 115(1), pp.347~354 (2016)[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]
- Trott O, "AutoDock Vina : improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading", J Comput Chem, 31(2), pp.455~461 (2010)[DOI]
- Won JY, "Functional characterization of Clonorchis sinensis choline transporter", Parasites Hosts Dis, 61(4), pp.428~438 (2023)[DOI]
- Cho SY, "Fifty years of the Korean society for parasitology", Korean J Parasitol, 47(Suppl), pp.S7~S19 (2009)[DOI]
- Chan JD, "Ca(2)(+)channels and praziquantel : a view from the free world", Parasitol Int, 62(6), pp.619~628 (2013)[DOI]
- Woelfle M, "Resolution of praziquantel", PLoS Negl Trop Dis, 5(9), pp.e1260~ (2011)[DOI]
- Doenhoff MJ, "Resistance of Schistosoma mansoni to praziquantel : is there a problem?", Trans R Soc Trop Med Hyg, 96(5), pp.465~469 (2002)[DOI]
- Joet T, "Validation of the hexose transporter of Plasmodium falciparum as a novel drug target", Proc Natl Acad Sci USA, 100(13), pp.7476~7479 (2003)[DOI]
- Saier MH Jr, "Families of transmembrane sugar transport proteins", Mol Microbiol, 35(4), pp.699~710 (2000)[DOI]
- Bueding E, "Carbohydrate metabolism of Schistosoma mansoni", J Gen Physiol, 33(5), pp.475~495 (1950)[DOI]
- Feistel T, "An expression system to screen for inhibitors of parasite glucose transporters", Mol Biochem Parasitol, 162(1), pp.71~76 (2008)[DOI]
- Githui EK, "Schistosoma mansoni : biochemical characterization of lactate transporters or similar proteins", Exp Parasitol, 114(3), pp.180~188 (2006)[DOI]
- Han SS, "The uptake of 14C-glucose by Clonorchis sinensis", Korean J Med, 4(4), pp.23~28 (1961)
- Burchmore RJ, "Genetic characterization of glucose transporter function in Leishmania mexicana", Proc Natl Acad Sci USA, 100(7), pp.3901~3906 (2003)[DOI]
- Dai F, "Glucose transporters and sodium glucose co-transporters cooperatively import glucose into energy-demanding organs in carcinogenic liver fluke Clonorchis sinensis", PLoS Negl Trop Dis, 18(7), pp.e0012315~ (2024)[DOI]
- Dai F, "Sodium-bile acid co-transporter is crucial for survival of a carcinogenic liver fluke Clonorchis sinensis in the bile", PLoS Negl Trop Dis, 14(12), pp.e0008952~ (2020)[DOI]
- Jung H, "Functional characterization of Clonorchis sinensis sodium-bile acid co-transporter(CsSBAT)as a steroid sulfate transporter", Parasitol Res, 121(1), pp.217~224 (2022)[DOI]