Abstract
MDCK cells expressing an inducible duodenal cytochrome b-green fluorescent protein (Dcytb-EGFP) fusion construct were used to investigate the function of Dcytb. The Dcytb-EGFP protein was targeted correctly to the plasma membrane, and cells displayed increased ferric and cupric reductase activities, which were greatly reduced in the presence of doxycycline. The data suggests that Dcytb plays a physiological role in both iron and copper uptake, through divalent metal transporter 1 (DMT1) and copper transporter 1, respectively. In support of this hypothesis, we show that 59Fe uptake was significantly enhanced in Dcytb-EGFP expressing MDCK cells which endogenously express DMT1.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Cation Transport Proteins / metabolism
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Cell Line
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Copper / metabolism*
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Copper Transporter 1
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Cytochrome b Group / genetics
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Cytochrome b Group / metabolism*
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Dogs
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FMN Reductase / genetics
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FMN Reductase / metabolism*
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Green Fluorescent Proteins / genetics
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Ion Transport
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Iron / metabolism*
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Mice
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Oxidoreductases / genetics
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Oxidoreductases / metabolism*
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Substrate Specificity
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Transfection
Substances
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Cation Transport Proteins
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Copper Transporter 1
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Cytochrome b Group
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Recombinant Fusion Proteins
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Slc31a1 protein, mouse
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enhanced green fluorescent protein
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solute carrier family 11- (proton-coupled divalent metal ion transporters), member 2
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Green Fluorescent Proteins
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Copper
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Iron
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Oxidoreductases
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cupric reductase (NADH-linked)
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FMN Reductase
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Cybrd1 protein, mouse
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ferric citrate iron reductase