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首页> 美国卫生研究院文献>Elsevier Public Health Emergency Collection >Establishment of Stable Cell Lines With High Expression of Heterodimers of Human 4F2hc and Human Amino Acid Transporter LAT1 or LAT2 and Delineation of Their Differential Interaction With α-Alkyl Moieties
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Establishment of Stable Cell Lines With High Expression of Heterodimers of Human 4F2hc and Human Amino Acid Transporter LAT1 or LAT2 and Delineation of Their Differential Interaction With α-Alkyl Moieties

机译：高表达人4F2hc和人氨基酸转运蛋白LAT1或LAT2异源二聚体的稳定细胞系的建立及其与α-烷基部分的微分相互作用的描述

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摘要

System L is a major transport system for cellular uptake of neutral amino acids. Among system L transporters, L-type amino acid transporter 1 (LAT1) is responsible for the nutrient uptake in cancer cells, whereas L-type amino acid transporter 2 (LAT2) is a transporter for non-cancer cells. In this study, we have established HEK293 cell lines stably expressing high levels of human LAT1 and LAT2 forming heterodimers with native human 4F2hc of the cells. We have found that -[ C]alanine is an appropriate substrate to examine the function of LAT2, whereas -[ C]leucine is used for LAT1. By using -[ C]alanine on LAT2, we have for the first time directly evaluated the function of human LAT2 expressed in mammalian cells and obtained its reliable kinetics. Using -alkyl amino acids including -methyl-alanine and -ethyl- -alanine, we have demonstrated that -alkyl groups interfere with the interaction with LAT2. These cell lines with higher practical advantages would be useful for screening and analyzing compounds to develop LAT1-specific drugs that can be used for cancer diagnosis and therapeutics. The strategy that we took to establish the cell lines would also be applicable to the other heterodimeric transporters with important therapeutic implications.
机译：系统L是细胞摄取中性氨基酸的主要转运系统。在系统L转运蛋白中，L型氨基酸转运蛋白1（LAT1）负责癌细胞中的营养摄取，而L型氨基酸转运蛋白2（LAT2）是非癌细胞的转运蛋白。在这项研究中，我们已经建立了HEK293细胞系，该细胞系可稳定表达高水平的人LAT1和LAT2，并与天然人4F2hc形成异源二聚体。我们发现-[C]丙氨酸是检查LAT2功能的合适底物，而-[C]亮氨酸用于LAT1。通过在LAT2上使用-[C]丙氨酸，我们首次直接评估了在哺乳动物细胞中表达的人LAT2的功能，并获得了其可靠的动力学。使用包括-甲基-丙氨酸和-乙基-丙氨酸的-烷基氨基酸，我们已经证明-烷基干扰与LAT2的相互作用。这些具有较高实际优势的细胞系可用于筛选和分析化合物，以开发可用于癌症诊断和治疗的LAT1特异性药物。我们用于建立细胞系的策略也将适用于具有重要治疗意义的其他异二聚体转运蛋白。

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