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Functional characterization of Clonorchis sinensis choline transporter

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...

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Autores principales: Won, Jeong Yeon, Louis, Johnsy Mary, Roh, Eui Sun, Cha, Seok Ho, Han, Jin-Hee
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Korean Society for Parasitology and Tropical Medicine 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10693965/
https://www.ncbi.nlm.nih.gov/pubmed/38043538
http://dx.doi.org/10.3347/PHD.23082
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author Won, Jeong Yeon
Louis, Johnsy Mary
Roh, Eui Sun
Cha, Seok Ho
Han, Jin-Hee
author_facet Won, Jeong Yeon
Louis, Johnsy Mary
Roh, Eui Sun
Cha, Seok Ho
Han, Jin-Hee
author_sort Won, Jeong Yeon
collection PubMed
description 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.
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spelling pubmed-106939652023-12-04 Functional characterization of Clonorchis sinensis choline transporter Won, Jeong Yeon Louis, Johnsy Mary Roh, Eui Sun Cha, Seok Ho Han, Jin-Hee Parasites Hosts Dis Original Article 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. The Korean Society for Parasitology and Tropical Medicine 2023-11 2023-11-28 /pmc/articles/PMC10693965/ /pubmed/38043538 http://dx.doi.org/10.3347/PHD.23082 Text en © 2023 The Korean Society for Parasitology and Tropical Medicine https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (https://creativecommons.org/licenses/by-nc/4.0 (https://creativecommons.org/licenses/by-nc/4.0/) ) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Article
Won, Jeong Yeon
Louis, Johnsy Mary
Roh, Eui Sun
Cha, Seok Ho
Han, Jin-Hee
Functional characterization of Clonorchis sinensis choline transporter
title Functional characterization of Clonorchis sinensis choline transporter
title_full Functional characterization of Clonorchis sinensis choline transporter
title_fullStr Functional characterization of Clonorchis sinensis choline transporter
title_full_unstemmed Functional characterization of Clonorchis sinensis choline transporter
title_short Functional characterization of Clonorchis sinensis choline transporter
title_sort functional characterization of clonorchis sinensis choline transporter
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10693965/
https://www.ncbi.nlm.nih.gov/pubmed/38043538
http://dx.doi.org/10.3347/PHD.23082
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