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Transporters of glycine, cystine, glutamate and glutamine in canine lens epithelial cells
Author(s) -
OCHIAI H
Publication year - 2013
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2013.s074.x
Subject(s) - glutamine , cystine , glycine , amino acid , amino acid transporter , transporter , microbiology and biotechnology , glutamate aspartate transporter , biochemistry , glutamate receptor , chemistry , western blot , biology , excitatory amino acid transporter , cysteine , enzyme , gene , receptor
Purpose The aim of this study is to evaluate GSH synthesis‐related amino acid (glycine, cystine, glutamate and glutamine) transport activity in a canine lens epithelial cell line (cdLEC) and define the transporter. Methods The primary lens epithelial cells of mature cataract dog were transfected with the expression plasmid DNA of large T antigen from replication origin‐defective simian virus 40 (SV40), then cloned using glass a cylinder. The transport activities of four amino acids in lens epithelial cells were determined. The cDNA sequences of transporter contributing GSH synthesis were also determined. Results Na‐dependent glycine and glutamate transport activity was 2.70 + 0.26 and 4.40 + 1.10 nmol/mg protein min, respectively. The cystine transport was Na‐independent and the transport activity was 0.626 + 0.17 nmol/mg protein min. The cDNA sequence of the glycine transporter 1(GlyT1), cystine/glutamate exchanger (xCT,), Na‐dependent neutral amino acid transporter 2(ASCT2), and glutamate transporter (EAAT) were determined. Western blot analysis confirmed their expression in membrane protein sample of cdLEC. Conclusion Lens epithelial cells possess GlyT1, xCT, ASCT2 and EAAT1‐5.