Mechanism of spermidine uptake in cultured mammalian cells and its inhibition by some polyamine analogues.
Transport pathways for spermidine (Spd) were characterized in mammalian cells in culture of different origin, i.e. L 1210, P 388, C 6, U 251, Balb/c 3T3 normal and transformed by virus SV40 (SV40/3T3). The kinetic constants (Km and Vmax) for 14C-Spd uptake were found to be different in these cells. Spd uptake was inhibited by spermine and putrescine in all cells. Preloading of these cells with system A and other amino acids, including ornithine, usually did not affect Spd uptake, except in L 1210 and C 6 cells, where Spd uptake was accelerated by 2-aminoisobutyric acid, demonstrating that in these two cell lines the polyamines share the system A pathway. Iso-osmotic replacement of Na+ by choline chloride in the assay medium resulted in a decrease in Spd uptake which suggests that Spd uptake is Na+ activated. In all cells, Spd uptake was inhibited by gramicidin and the Ca2+ ionophore A 23187. The degree of inhibition varied among the cells. Valinomycin (K+ ionophore) inhibited Spd uptake by C 6, P 388, Balb/c 3T3 and SV40/3T3 but not by L 1210 and U 251 cells. Treatment with N-ethylmaleimide or p-L 1210, C 6, Balb/c 3T3 and SV40/3T3 cells did not affect appreciably the uptake process. Some newly synthesized polyamine analogues inhibited the Spd uptake of all cells.
Khan
N A
NA
Department of Cell Biology, Faculty of Medicine, Central Hospital of the University of Rennes-I, France.
Quemener
V
V
Seiler
N
N
Moulinoux
J P
JP
D000596:Amino Acids / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D000001:Calcimycin / Q000494:pharmacology; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology*; D005910:Glioma / Q000378:metabolism / Q000473:pathology*; D006096:Gramicidin / Q000494:pharmacology; D006801:Humans; D007939:Leukemia L1210 / Q000378:metabolism / Q000473:pathology*; D007941:Leukemia P388 / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008807:Mice, Inbred BALB C; D011073:Polyamines / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D012964:Sodium / Q000494:pharmacology; D013095:Spermidine / Q000493:pharmacokinetics*; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000473:pathology / Q000502:physiology; D014634:Valinomycin / Q000494:pharmacology
D000596:Amino Acids; D011073:Polyamines; D013439:Sulfhydryl Reagents; D006096:Gramicidin; D014634:Valinomycin; D000001:Calcimycin; D012964:Sodium; D013095:Spermidine