Characterization of polyamine transport in rat aortic smooth muscle cells.
Increased arterial wall polyamine content has been linked to intimal hyperplasia (IH) formation. Intracellular polyamine content may be regulated by a polyamine transmembrane transport mechanism, but the existence of such a system has not been demonstrated in systemic arterial smooth muscle cells. This study characterizes polyamine transport as found in rat aortic smooth muscle cells. Smooth muscle cells were isolated and cultured from Sprague-Dawley rat aortas. Polyamine transport was determined by adding [14C]-polyamines to the medium, calculating transport kinetic parameters, Vmax and Km. Competition studies with unlabeled polyamines and uptake in the presence of paraquat, a polyamine transport inhibitor, were done to test the specificity of the uptake system. We identified polyamine transporters in aortic smooth muscle cells which were temperature, concentration, and time dependent. Kinetic studies revealed that spermidine and spermine had greater affinity for the transporter(s) than putrescine (Km = 0.3, 0.3, and 3.7 microM respectively; P = 0.0001) while maximum uptake velocity was similar for all polyamines (26.6-31.0 pmole/mg protein/min). Inhibition of de novo polyamine synthesis upregulated polyamine transport 2.8-3.8 times (P = 0.0001) while transporter affinity (as reflected by Km) remained unchanged. Competition studies and paraquat treatment indicated the presence of two polyamine transporters: one shared by all polyamines, the other specific for spermine and spermidine. These data indicate that transmembrane polyamine transport occurs in arterial smooth muscle cells. Upregulation of this system may represent one control mechanism for IH development.
Toursarkissian
B
B
Department of Surgery, College of Medicine, University of Kentucky Medical Center, Lexington 40536-0084.
Endean
E D
ED
Aziz
S M
SM
D000818:Animals; D001013:Aorta, Thoracic / Q000378:metabolism; D001667:Binding, Competitive; D001692:Biological Transport / Q000187:drug effects; D002250:Carbon Radioisotopes; D002478:Cells, Cultured; D000518:Eflornithine / Q000494:pharmacology; D007700:Kinetics; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D010269:Paraquat / Q000494:pharmacology; D011073:Polyamines / Q000378:metabolism*; D011700:Putrescine / Q000378:metabolism / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013095:Spermidine / Q000378:metabolism / Q000494:pharmacology; D013096:Spermine / Q000378:metabolism / Q000494:pharmacology
D002250:Carbon Radioisotopes; D011073:Polyamines; D013096:Spermine; D010269:Paraquat; D013095:Spermidine; D011700:Putrescine; D000518:Eflornithine