Characterization of L-glutamine transport by a human neuroblastoma cell line.

摘要

This study characterized the Na+-dependent transport of L-glutamine by a human neuroblastoma cell line, SK-N-SH. The Na+-dependent component represented >95% of the total glutamine uptake. Kinetic studies showed a single saturable high-affinity carrier with a Michaelis constant (K(m)) of 163 +/- 23 microM and a maximum transport velocity (Vmax) of 13,713 +/- 803 pmol x mg protein(-1) x min(-1). Glutamine uptake was markedly inhibited in the presence of L-alanine, L-asparagine, and L-serine. Li+ did not substitute for Na+. These data show that L-glutamine is predominantly taken up through system ASC. Glutamine deprivation resulted in the decrease of glutamine transport by a mechanism that decreased Vmax without affecting K(m). The expression of the system ASC subtype ASCT2 decreased in the glutamine-deprived group, whereas glutamine deprivation did not induce changes in system ASC subtype ASCT1 mRNA expression. Adaptive increases in Na+-dependent glutamate, Na+-dependent 2-(methylamino)isobutyric acid, and Na+-independent leucine transport were observed under glutamine-deprived conditions, which were completely blocked by actinomycin D and cycloheximide. These mechanisms may allow cells to survive and even grow under nutrient-deprived conditions.