Specificities of red cell membrane sites transporting three long chain fatty acids.
We studied the specificities of human red cell membrane bindings of three long chain fatty acids, palmitic- arachidonic- and oleic acid, using resealed membranes, ghosts. Previously estimated binding capacities, affinities and inside/outside distributions suggest separated binding sites. This possibility is explored by estimating the binding properties of one fatty acid in the presence of one or two of the others. Binding capacities, nmol g-1 ghosts, of palmitic and arachidonic acid estimated simultaneously vs. separately are 27.4 +/- 2.7 vs. 29.0 +/- 2.1 (P < 0.6) and 6.5 +/- 0.6 vs. 5.5 +/- 0.5 (P < 0.2) respectively. The corresponding estimates for oleic- and palmitic acid are 36.5 +/- 2.0 vs. 34.0 +/- 2.2 (P < 0.4) and 28.4 +/- 1.8 versus 29.1 +/- 2.1 (P < 0.8). The binding sites are therefore independent. For each of the three fatty acids in the absence or in the presence of one or two of the others, the inside/outside distributions of the binding sites and the membrane transfer rate constants are elucidated by exchange efflux kinetics at 0 degrees C from ghosts with and without enclosed albumin. Packed ghosts loaded with radioactive acids are injected rapidly into a large volume of vigorously stirred buffer with albumin. With a resolution time of about 1-sec serial filtered ghost-free aliquots are collected and counted. The analyses show that palmitic- and oleic acid sites of transport are entirely independent but do not exclude that palmitic- and/or oleic acid binding may diminish the arachidonic acid affinity a little. The diversity combined with specificity suggests that the transport sites for long chain fatty acids are protein-determined microdomains of phospholipids.
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Department of Medical Biochemistry and Genetics, University of Copenhagen, Panum Institute, Denmark.
Bojesen
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D001692:Biological Transport; D004910:Erythrocyte Membrane / Q000378:metabolism*; D005227:Fatty Acids / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008432:Mathematical Computing; D008954:Models, Biological; D012710:Serum Albumin, Bovine
D005227:Fatty Acids; D012710:Serum Albumin, Bovine