Saturable stimulation of fatty acid transport through model cytoplasm by soluble binding protein.
To better define the role of soluble binding proteins in the cytoplasmic transport of amphipathic molecules, we measured the diffusional mobility of a fluorescent long-chain fatty acid, 12-N-methyl-(7-nitrobenz-2-oxa-1,3-diazol)aminostearate (NBD-stearate), through model cytoplasm as a function of soluble binding protein concentration. Diffusional mobilities were correlated with the partition of the fatty acid between membrane and protein binding sites. Cytoplasm was modeled as a dense suspension of liposomes, and albumin was used as a model binding protein. Albumin saturably increased NBD-stearate mobility through the membrane suspension approximately eightfold. Fatty acid mobility in the absence of albumin was identical to the mobility of the membrane vesicles (1.99 +/- 0.33 x 10(-8) cm(2)/s), whereas the mobility at saturating concentrations was identical to the mobility of albumin (1.65 +/- 0.12 x 10(-7) cm(2)/s). The protein concentration producing half-maximal stimulation of NBD-stearate diffusion (42.8 +/- 0.3 microM) was unexpectedly greater than that required to solubilize half of the NBD-stearate (17.9 +/- 3.0 microM). These results support a proposed mechanism for cytoplasmic transport of small amphipathic molecules in which aqueous diffusion of the protein-bound form of the molecule largely determines the transport rate. However, slow interchange of fatty acid between the binding protein and membranes also appears to influence the transport rate in this model system.
Department of Medicine and the Liver Center, University of California, San Francisco, California 94143-0538, USA. dickw@itsa.ucsf.edu
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D009327:4-Chloro-7-nitrobenzofurazan / Q000031:analogs & derivatives / Q000378:metabolism; D000418:Albumins / Q000378:metabolism / Q000494:pharmacology; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D002498:Centrifugation; D005591:Chemical Fractionation; D003593:Cytoplasm / Q000378:metabolism*; D004058:Diffusion; D005227:Fatty Acids / Q000378:metabolism*; D008081:Liposomes / Q000378:metabolism* / Q000648:ultrastructure; D008954:Models, Biological; D009994:Osmolar Concentration; D012710:Serum Albumin, Bovine / Q000378:metabolism / Q000494:pharmacology; D013228:Stearates / Q000378:metabolism
D000418:Albumins; D002352:Carrier Proteins; D005227:Fatty Acids; D008081:Liposomes; D013228:Stearates; C060027:12-N-methyl-7-nitrobenzo-2-oxa-1,3-diazolamino stearate; D012710:Serum Albumin, Bovine; D009327:4-Chloro-7-nitrobenzofurazan