Document Detail


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.
MedLine Citation:
PMID:  22850614     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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 L-[(14)C]alanine is an appropriate substrate to examine the function of LAT2, whereas L-[(14)C]leucine is used for LAT1. By using L-[(14)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 α-alkyl amino acids including α-methyl-alanine and α-ethyl-L-alanine, we have demonstrated that α-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.
Authors:
Narakorn Khunweeraphong; Shushi Nagamori; Pattama Wiriyasermkul; Yumiko Nishinaka; Printip Wongthai; Ryuichi Ohgaki; Hidekazu Tanaka; Hideyuki Tominaga; Hiroyuki Sakurai; Yoshikatsu Kanai
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-07-31
Journal Detail:
Title:  Journal of pharmacological sciences     Volume:  119     ISSN:  1347-8648     ISO Abbreviation:  J. Pharmacol. Sci.     Publication Date:  2012  
Date Detail:
Created Date:  2012-09-14     Completed Date:  2013-01-29     Revised Date:  2013-06-06    
Medline Journal Info:
Nlm Unique ID:  101167001     Medline TA:  J Pharmacol Sci     Country:  Japan    
Other Details:
Languages:  eng     Pagination:  368-80     Citation Subset:  IM    
Affiliation:
Division of Bio-system Pharmacology, Department of Pharmacology, Graduate School of Medicine, Osaka University, Japan.
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MeSH Terms
Descriptor/Qualifier:
Adaptor Proteins, Signal Transducing / chemistry,  genetics*,  metabolism
Amino Acids / pharmacology
Antigens, CD98 Heavy Chain / chemistry,  genetics*,  metabolism
Biological Transport / drug effects
HEK293 Cells / metabolism*
Humans
Large Neutral Amino Acid-Transporter 1 / chemistry,  genetics*,  metabolism
Protein Multimerization
Chemical
Reg. No./Substance:
0/Adaptor Proteins, Signal Transducing; 0/Amino Acids; 0/Antigens, CD98 Heavy Chain; 0/LAT2 protein, human; 0/Large Neutral Amino Acid-Transporter 1; 0/SLC3A2 protein, human

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