Analysis and generation of functional molecules to modulate neutral amino acid transporters

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Alanine-serine-cysteine transporters (ASCT1, 2) are neutral amino acids transporters. Their representative substrates are alanine, serine and cysteine. Since th ere is no specific inhibitors or substrates for ASCT1 and ASCT2, their physiological functions are quite mystery. To develop their specific compounds to modulate their functions, high-throughput screening is essential. Therefore, we made two approaches as follows. First, we looked for host cells with lower level of endogenous ASCT1 or ASCT2. We checked CHO cells, HEK293 cells, 3T3 cells and other cells. But all of the cells that we examined showed higher level of ASCTs expression. Second, we tried the cloning of human and rat type ASCTs. Cloned ASCTs were integrated into the expression vectors for their expressions in mammalian cells and Xenopus oocytes.