Characterization of several amino acid transports and glutamine metabolism in MOLT4 human T4 leukemia cells.
The transport system responsible for glutamine, alanine and glutamate in MOLT4 human T4 leukemia cell line were characterized. Kinetic studies of sodium-dependent glutamine and alanine transport exhibited a single saturable high-affinity carrier with a Michaelis constant of 152 +/- 26 microm and 203 +/- 36 microm and a maximal transport velocity of 960 +/- 165 and 1096 +/- 208 nmol/10(9)cells/min, respectively. Glutamate uptake was less than one-tenth of glutamine and alanine, and linearly increased with glutamate concentration which was mediated by diffusion. 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid (SITS), known as anion channel blockers, inhibited the sodium-dependent glutamine and alanine transport by 40% at 10 microm. Cellular contents of these amino acids in MOLT4 cells revealed glutamate to be the highest among them despite low glutamate influx. A glutamine metabolism study using whole cells indicated this high conversion rate from glutamine to glutamate, but no conversion to another amino acid. Based on these results, the high glutamate concentration in MOLT4 was speculated to be synthesized from transported glutamine by active glutaminase.
Ochiai
H
H
Research Institute of Biosciences, School of Veterinary Medicine, Azabu University, 1-17-71 Fuchinobe, Sagamihara, Kanagawa 229-8501, Japan. ochiaih@azabu-u.ac.jp
Higa
K
K
Hishiyama
N
N
Hisamatsu
S
S
Fujise
H
H
D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid / Q000494:pharmacology; D000409:Alanine / Q000378:metabolism*; D026962:Amino Acid Transport System ASC / Q000378:metabolism; D001692:Biological Transport / Q000502:physiology; D015496:CD4-Positive T-Lymphocytes; D045744:Cell Line, Tumor / Q000378:metabolism; D005972:Glutaminase / Q000378:metabolism; D005973:Glutamine / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D007945:Leukemia, Lymphoid / Q000201:enzymology / Q000378:metabolism*
D026962:Amino Acid Transport System ASC; D005973:Glutamine; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid; D005972:Glutaminase; D000409:Alanine