Plasma membrane transport of 2-ketoisocaproate by rat hepatocytes in primary culture.
Others have shown that the branched chain 2-keto acids are generated in muscle, released into the bloodstream, and then removed by the liver where further catabolism occurs. The present investigation describes the plasma membrane transport systems for these metabolites in cultured rat hepatocytes. One of these systems in Na+-dependent, concentrates the 2-keto acids against a gradient, and is inhibited by pyruvate. The second process is Na+-independent, is less concentrative, and may be composed of two distinct systems as suggested by pyruvate inhibition studies. None of these systems accept neutral amino acids. For the transport of 2-ketoisocaproate, the Na+-dependent system exhibits a Km value of about 5 mM, whereas the corresponding value for the Na+-independent agency is 60 microM. The activity of the Na+-dependent system is moderately increased by insulin treatment of the cells, while neither agency is stimulated by glucagon, dexamethasone, or the combination of these two hormones. Hepatocytes from diabetic rats show enhanced transport by the Na+-dependent system and incubation of cultured hepatocytes for 24 h in the absence of 2-keto acids results in a 3-fold stimulation of the Na+-dependent system, but has no effect on the rate of Na+-independent transport. These results demonstrate the existence of at least two saturable transport systems for the branched chain 2-keto acids in the rat hepatocyte and the ability of the Na+-dependent system to respond to the extracellular environment.
D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D003907:Dexamethasone / Q000494:pharmacology; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism; D005934:Glucagon / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D007651:Keto Acids / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000378:metabolism*; D008297:Male; D011773:Pyruvates / Q000494:pharmacology; D019289:Pyruvic Acid; D051381:Rats; D011919:Rats, Inbred Strains; D012964:Sodium / Q000494:pharmacology
D007328:Insulin; D007651:Keto Acids; D011773:Pyruvates; D003907:Dexamethasone; C013082:alpha-ketoisocaproic acid; D019289:Pyruvic Acid; D005934:Glucagon; D012964:Sodium