Protein kinase C-dependent regulation of L-arginine transport activity in Caco-2 intestinal cells.
The regulation of plasma membrane L-arginine transport activity was investigated in differentiated and undifferentiated states of the human intestinal cell line, Caco-2. The sodium-independent, leucine-insensitive uptake of L-arginine measured in this study has been assigned by us previously to system y+ in Caco-2 cells. Treatment of cells with serum-free media containing epidermal growth factor (EGF), transforming growth factor alpha (TGF alpha), or the protein kinase C (PKC) activator 12-O-tetradecanoylphorbol 13-acetate (TPA), stimulated system y+ arginine transport activity in Caco-2 cells. Transport upregulation by these growth factors or by TPA was blocked by cycloheximide or the PKC inhibitor chelerythrine. Arginine uptake was diminished during the course of differentiation, attributable to a reduction in the transport system y+ capacity (Vmax) with no change in apparent affinity (Km). TPA stimulated arginine uptake required at least 3 h of continual exposure, and increased the membrane's transport capacity (Vmax) in both undifferentiated and differentiated cells. TPA elevated the diminished transport Vmax of differentiated cells TPA to the elevated Vmax value associated with undifferentiated cells. We conclude that upregulation of arginine transport is part of a pleiotropic response to EGF/TGF alpha, and that this involves PKC and de novo synthesis of polypeptides associated with system y+ transport activity.
Pan
M
M
Department of Physiology, College of Medicine, University of Florida, Gainesville 32610-0274, USA.
Stevens
B R
BR
D000470:Alkaloids; D001120:Arginine / Q000378:metabolism*; D053119:Benzophenanthridines; D001692:Biological Transport / Q000187:drug effects; D018938:Caco-2 Cells; D002352:Carrier Proteins / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects; D003513:Cycloheximide / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004815:Epidermal Growth Factor / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000378:metabolism*; D010617:Phenanthridines / Q000494:pharmacology; D011493:Protein Kinase C / Q000819:agonists / Q000037:antagonists & inhibitors / Q000502:physiology*; D011991:Receptors, Virus; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D016211:Transforming Growth Factor alpha / Q000494:pharmacology; D015854:Up-Regulation
D000470:Alkaloids; D053119:Benzophenanthridines; D002352:Carrier Proteins; D004791:Enzyme Inhibitors; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D010617:Phenanthridines; D011991:Receptors, Virus; D016211:Transforming Growth Factor alpha; C070506:ecotropic murine leukemia virus receptor; D004815:Epidermal Growth Factor; D001120:Arginine; D003513:Cycloheximide; C016299:chelerythrine; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate