Nucleoside transport in mammalian cell membranes. III. Kinetic and chemical modification studies of cytosine-arabinoside and uridine transport in hamster cells in culture.
Transport of the nucleoside analog cytosine-arabinoside (CAR) in transformed hamster cells in culture has been studied in conditions of minimal metabolic conversion. Uptake (zero-trans in) properties at 20 degrees C over a limited range of CAR concentrations were characterized by a Km of 350 micrometer and a maximal velocity (V) of 780 micrometer.min-1 (V/Km = 2.28 min-1). Equilibrium exhcange at 20 degrees C over a wider range of concentrations was best described by a saturable component with a Km of 500 micrometer and a v of 1230 micrometer.min-1 (V/Km = 2.26 min-1) and either a saturable component of high Km or a nonsaturable component of k = 0.3 min-1. For the saturable component, the v/Km values were similar in both procedures. CAR transport was inhibited by various metabolizable nucleosides. Uptake of some of these nucleosides was inhibited by CAR. CAR transport and uridine uptake were inhibited in a reversible but partially competitive fashion by high affinity probes like S-(p-nitrobenzyl-6-mercaptoinosine (NBMI) (Ki less than 0.5 nM) and in an irreversible fashion by SH reagents such as N-ethylmaleiimide (NEM). The organomercurial p-hydroxymercuribenzene sulfonate (pMBS) markedly stimulated transport of these nucleosides, but also markedly potentiated the inhibitory effects of either NBMI or NEM. The effects are interpreted either in terms of models which invoke allosteric properties or in terms of two transport systems which display distinct chemical susceptibilities to externally added probes.
D000345:Affinity Labels; D001693:Biological Transport, Active / Q000187:drug effects; D002460:Cell Line; D003561:Cytarabine / Q000378:metabolism*; D005033:Ethylmaleimide / Q000494:pharmacology; D007700:Kinetics; D010663:Phenylmercury Compounds / Q000494:pharmacology; D013868:Thioinosine / Q000031:analogs & derivatives / Q000494:pharmacology; D014529:Uridine / Q000378:metabolism*
D000345:Affinity Labels; D010663:Phenylmercury Compounds; D003561:Cytarabine; D013868:Thioinosine; D005033:Ethylmaleimide; D014529:Uridine