title: Iron binding to, and release from, the basolateral membrane of mouse duodenal enterocytes.
abstract: The basolateral membrane of mouse duodenal enterocytes can be selectively labelled in vitro with 59Fe by incubating intact enterocytes with 59Fe(III)-nitrilotriacetate at 0-4 degrees C. It has been proposed that this labelling represents binding to a site important in the transfer of intracellular Fe to the portal plasma (Snape, S., Simpson, R.J. and Peters, T.J. (1990) Cell Biochem. Funct. 8, 107-115). Studies presented here show binding to intact enterocytes in vitro was complete within 1 h and was proportional to enterocyte protein concentration. Binding to enterocytes isolated from both normal and chronically hypoxic mice showed a hyperbolic dependence on medium Fe(III) concentration, consistent with a single class of binding sites. Neither apparent binding constant nor maximal binding were increased by hypoxic exposure of mice, suggesting that the increased in vivo labelling of this site in hypoxia is not due to an increase in affinity or capacity of this site for iron. Release of iron from intact enterocytes, labelled at 0-4 degrees C, was measured at 37 degrees C and 0-4 degrees C. Release of 59Fe was extensive and more rapid at 37 degrees C with highest release to mouse serum. Iron released to serum was found to be bound to transferrin. Prior dialysis of serum against buffer led to complete failure of enterocytes to release iron. Reconstituting serum by adding back the dialysate restores release to levels seen in fresh serum, suggesting that low molecular weight serum components, notably bicarbonate, mediate iron transfer from the basolateral membrane to serum transferrin. The properties of the basolateral membrane iron binding site described here are consistent with a role in the iron transfer process.
authors.affiliation: Department Clinical Biochemistry, Kings College School of Medicine and Dentistry, London, U.K.
authors.forename: S
authors.initials: S
authors.lastname: Snape
authors.forename: R J
authors.initials: RJ
authors.lastname: Simpson
mesh_terms: D000818:Animals; D001798:Blood Proteins / Q000378:metabolism; D002458:Cell Fractionation; D002462:Cell Membrane / Q000378:metabolism; D002614:Chelating Agents / Q000378:metabolism; D004386:Duodenum / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D007501:Iron / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D008670:Metals / Q000494:pharmacology; D051379:Mice; D008970:Molecular Weight; D014168:Transferrin / Q000378:metabolism
chemical_list: D001798:Blood Proteins; D002614:Chelating Agents; D008670:Metals; D014168:Transferrin; D007501:Iron