Cargando…
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...
Autores principales: | , , , , |
---|---|
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 |
_version_ | 1785153272971001856 |
---|---|
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. |
format | Online Article Text |
id | pubmed-10693965 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | The Korean Society for Parasitology and Tropical Medicine |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT wonjeongyeon functionalcharacterizationofclonorchissinensischolinetransporter AT louisjohnsymary functionalcharacterizationofclonorchissinensischolinetransporter AT roheuisun functionalcharacterizationofclonorchissinensischolinetransporter AT chaseokho functionalcharacterizationofclonorchissinensischolinetransporter AT hanjinhee functionalcharacterizationofclonorchissinensischolinetransporter |