A rapid and simple non-radioactive assay for measuring uptake by solute carrier transporters
作者全名：Song, Kunling; Zhang, Longbin; Fu, Xian; Li, Linfeng; Zhu, Gaolin; Wu, Mingjun; Zhang, Wei; He, Jia; Zhu, Sanyong; Dang, Yongjun; Liu, Jun-Yan; Chen, Chang; Guo, Zufeng
作者地址：[Song, Kunling; Zhang, Longbin; Fu, Xian; Li, Linfeng; Zhu, Gaolin; Zhang, Wei; He, Jia; Zhu, Sanyong; Dang, Yongjun; Liu, Jun-Yan; Guo, Zufeng] Chongqing Med Univ, Affiliated Hosp 2, Inst Life Sci, Basic Med Res & Innovat Ctr Novel Target & Therape, Chongqing, Peoples R China; [Song, Kunling; Zhang, Longbin; Fu, Xian; Li, Linfeng; Zhu, Gaolin; Zhang, Wei; He, Jia; Zhu, Sanyong; Dang, Yongjun; Liu, Jun-Yan; Guo, Zufeng] Chongqing Med Univ, Affiliated Hosp 2, Dept Anesthesiol, Chongqing, Peoples R China; [Wu, Mingjun; Chen, Chang] Chongqing Med Univ, Inst Life Sci, Chongqing, Peoples R China; [Dang, Yongjun; Liu, Jun-Yan; Guo, Zufeng] Chongqing Med Univ, Coll Pharm, Chongqing, Peoples R China
通信作者：Liu, JY; Guo, ZF (通讯作者)，Chongqing Med Univ, Affiliated Hosp 2, Inst Life Sci, Basic Med Res & Innovat Ctr Novel Target & Therape, Chongqing, Peoples R China.; Liu, JY; Guo, ZF (通讯作者)，Chongqing Med Univ, Affiliated Hosp 2, Dept Anesthesiol, Chongqing, Peoples R China.; Chen, C (通讯作者)，Chongqing Med Univ, Inst Life Sci, Chongqing, Peoples R China.; Liu, JY; Guo, ZF (通讯作者)，Chongqing Med Univ, Coll Pharm, Chongqing, Peoples R China.
来源：FRONTIERS IN PHARMACOLOGY
ESI学科分类：PHARMACOLOGY & TOXICOLOGY
WOS号：WOS:001215231300001
JCR分区：Q1
影响因子：4.4
年份：2024
卷号：15
期号：
开始页：
结束页：
文献类型：Article
关键词：solute carrier transporter; uptake assay; LC-MS/MS; LAT1; NTCP
摘要：Introduction: Solute carrier (SLC) transport proteins play a crucial role in maintaining cellular nutrient and metabolite homeostasis and are implicated in various human diseases, making them potential targets for therapeutic interventions. However, the study of SLCs has been limited due to the lack of suitable tools, particularly cell-based substrate uptake assays, necessary for understanding their biological functions and for drug discovery purposes.Methods: In this study, a cell-based uptake assay was developed using a stable isotope-labeled compound as the substrate for SLCs, with detection facilitated by liquid chromatography-tandem mass spectrometry (LC-MS/MS). This assay aimed to address the limitations of existing assays, such as reliance on hazardous radiolabeled substrates and limited availability of fluorescent biosensors.Results: The developed assay was successfully applied to detect substrate uptakes by two specific SLCs: L-type amino acid transporter 1 (LAT1) and sodium taurocholate co-transporting polypeptide (NTCP). Importantly, the assay demonstrated comparable results to the radioactive method, indicating its reliability and accuracy. Furthermore, the assay was utilized to screen for novel inhibitors of NTCP, leading to the identification of a potential NTCP inhibitor compound.Discussion: The findings highlight the utility of the developed cell-based uptake assay as a rapid, simple, and environmentally friendly tool for investigating SLCs' biological roles and for drug discovery purposes. This assay offers a safer alternative to traditional methods and has the potential to contribute significantly to advancing our understanding of SLC function and identifying therapeutic agents targeting SLC-mediated pathways.
基金机构：National Natural Science Foundation of China [22277010, 82071671]; Ministry of Human Resources and Social Security Funding Scheme for High-Level Overseas Chinese Students' Return of China; Natural Science Foundation of Chongqing [CSTB2022NSCQ-MSX1061]; Chongqing Human Resources and Social Security Bureau Innovation Funding Scheme for Overseas Chinese Students' Return; CQMU Program for Youth Innovation in Future Medicine [W0074]; High-Level Innovation Platform Cultivation Plan of Chongqing; University Innovation Research Group in Chongqing [CXQT21016]
基金资助正文：The author(s) declare that financial support was received for the research, authorship, and/or publication of this article. This work was supported by the National Natural Science Foundation of China [No. 22277010 to ZG and 82071671 to CC], Ministry of Human Resources and Social Security Funding Scheme for High-Level Overseas Chinese Students' Return of China [to ZG], Natural Science Foundation of Chongqing [No. CSTB2022NSCQ-MSX1061 to ZG], Chongqing Human Resources and Social Security Bureau Innovation Funding Scheme for Overseas Chinese Students' Return (to ZG), CQMU Program for Youth Innovation in Future Medicine [No. W0074 to ZG], High-Level Innovation Platform Cultivation Plan of Chongqing [to ZG, J-YL and YD), and University Innovation Research Group in Chongqing [No. CXQT21016 to ZG, J-YL, and YD].
