title: Zip14 is a complex broad-scope metal-ion transporter whose functional properties support roles in the cellular uptake of zinc and nontransferrin-bound iron.
abstract: Recent studies have shown that overexpression of the transmembrane protein Zrt- and Irt-like protein 14 (Zip14) stimulates the cellular uptake of zinc and nontransferrin-bound iron (NTBI). Here, we directly tested the hypothesis that Zip14 transports free zinc, iron, and other metal ions by using the Xenopus laevis oocyte heterologous expression system, and use of this approach also allowed us to characterize the functional properties of Zip14. Expression of mouse Zip14 in RNA-injected oocytes stimulated the uptake of (55)Fe in the presence of l-ascorbate but not nitrilotriacetic acid, indicating that Zip14 is an iron transporter specific for ferrous ion (Fe(2+)) over ferric ion (Fe(3+)). Zip14-mediated (55)Fe(2+) uptake was saturable (K(0.5) ≈ 2 μM), temperature-dependent (apparent activation energy, E(a) = 15 kcal/mol), pH-sensitive, Ca(2+)-dependent, and inhibited by Co(2+), Mn(2+), and Zn(2+). HCO(3)(-) stimulated (55)Fe(2+) transport. These properties are in close agreement with those of NTBI uptake in the perfused rat liver and in isolated hepatocytes reported in the literature. Zip14 also mediated the uptake of (109)Cd(2+), (54)Mn(2+), and (65)Zn(2+) but not (64)Cu (I or II). (65)Zn(2+) uptake also was saturable (K(0.5) ≈ 2 μM) but, notably, the metal-ion inhibition profile and Ca(2+) dependence of Zn(2+) transport differed from those of Fe(2+) transport, and we propose a model to account for these observations. Our data reveal that Zip14 is a complex, broad-scope metal-ion transporter. Whereas zinc appears to be a preferred substrate under normal conditions, we found that Zip14 is capable of mediating cellular uptake of NTBI characteristic of iron-overload conditions.
authors.affiliation: Department of Molecular and Cellular Physiology, University of Cincinnati College of Medicine, Ohio 45267-0576, USA.
authors.forename: Jorge J
authors.initials: JJ
authors.lastname: Pinilla-Tenas
authors.forename: Brian K
authors.initials: BK
authors.lastname: Sparkman
authors.forename: Ali
authors.initials: A
authors.lastname: Shawki
authors.forename: Anthony C
authors.initials: AC
authors.lastname: Illing
authors.forename: Colin J
authors.initials: CJ
authors.lastname: Mitchell
authors.forename: Ningning
authors.initials: N
authors.lastname: Zhao
authors.forename: Juan P
authors.initials: JP
authors.lastname: Liuzzi
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Cousins
authors.forename: Mitchell D
authors.initials: MD
authors.lastname: Knutson
authors.forename: Bryan
authors.initials: B
authors.lastname: Mackenzie
mesh_terms: D000818:Animals; D027682:Cation Transport Proteins / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation; D006801:Humans; D007501:Iron / Q000378:metabolism*; D051379:Mice; D009865:Oocytes; D020033:Protein Isoforms; D051381:Rats; D014981:Xenopus; D015032:Zinc / Q000378:metabolism*; D000099236:Solute Carrier Family 11, Member 2
chemical_list: D027682:Cation Transport Proteins; D007501:Iron; D020033:Protein Isoforms; D015032:Zinc; D000099236:Solute Carrier Family 11, Member 2; C500556:SLC39A14 protein, mouse