title: Investigation of the biophysical and cell biological properties of ferroportin, a multipass integral membrane protein iron exporter.
abstract: Ferroportin is a multipass membrane protein that serves as an iron exporter in many vertebrate cell types. Ferroportin-mediated iron export is controlled by the hormone hepcidin, which binds ferroportin, causing its internalization and degradation. Mutations in ferroportin cause a form of the iron overload hereditary disease hemochromatosis. Relatively little is known about ferroportin's properties or the mechanism by which mutations cause disease. In this study, we expressed and purified human ferroportin to characterize its biochemical/biophysical properties in solution and conducted cell biological studies in mammalian cells. We found that purified detergent-solubilized ferroportin is a well-folded monomer that binds hepcidin. In cell membranes, the N- and C-termini were both cytosolic, implying an even number of transmembrane regions, and ferroportin was mainly localized to the plasma membrane. Hepcidin addition resulted in a redistribution of ferroportin to intracellular compartments that labeled with early endosomal and lysosomal, but not Golgi, markers and that trafficked along microtubules. An analysis of 16 disease-related ferroportin mutants revealed that all were expressed and trafficked to the plasma membrane but that some were resistant to hepcidin-induced internalization. The characterizations reported here form a basis upon which models for ferroportin's role in regulating iron homeostasis in health and disease can be interpreted.
authors.affiliation: Graduate Option in Biochemistry and Molecular Biophysics, California Institute of Technology, Pasadena, CA 91125, USA.
authors.forename: Adrian E
authors.initials: AE
authors.lastname: Rice
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Mendez
authors.forename: Craig A
authors.initials: CA
authors.lastname: Hokanson
authors.forename: Douglas C
authors.initials: DC
authors.lastname: Rees
authors.forename: Pamela J
authors.initials: PJ
authors.lastname: Björkman
mesh_terms: D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000378:metabolism*; D027682:Cation Transport Proteins / Q000737:chemistry* / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry; D011992:Endosomes / Q000737:chemistry; D064451:Hepcidins; D006801:Humans; D008247:Lysosomes / Q000737:chemistry; D008954:Models, Biological; D008958:Models, Molecular; D008970:Molecular Weight; D050505:Mutant Proteins / Q000378:metabolism; D011485:Protein Binding; D021381:Protein Transport; D000098742:Ferroportin
chemical_list: D023181:Antimicrobial Cationic Peptides; D027682:Cation Transport Proteins; C578702:HAMP protein, human; D064451:Hepcidins; D050505:Mutant Proteins; D000098742:Ferroportin