PMID: 9528678Apr 7, 1998Paper

Specific binding to plasma membrane is the first step in the uptake of non-transferrin iron by cultured cells

Biochimica Et Biophysica Acta
J MusílkováJ Kovár

Abstract

We studied transport of non-transferrin iron into HeLa cells adapted for growth in defined medium, containing either 5 micrograms/ml of iron-saturated transferrin (HeLa/Tf cells) or 5 microM ferric citrate (HeLa/Fe5 cells) as a source of iron. Employing 55Fe-ferric citrate, iron uptake by intact cells was compared with iron binding to isolated membranes. Uptake characteristics of both HeLa/Tf and HeLa/Fe5 cells seemed to be similar: Km = 14 microM and Vmax = 135 pmol Fe/min/10(5) cells for HeLa/Tf, Km = 22 microM and Vmax = 165 pmol Fe/min/10(5) cells for HeLa/Fe5. Increasing concentrations (0.3-1.2 microM) of 55Fe-ferric citrate, producing levels of free 55Fe which were independent of total Fe under the experimental conditions used, led to increased binding of 55Fe for both HeLa/Tf and HeLa/Fe5 cells (1.08-8.03 nmol Fe/h/10(5) cells). This corresponds with the suggestion that iron was bound in the form of ferric citrate rather than in the form of free iron. Dissociation constants of Fe binding, KD = 0.61 microM for HeLa/Tf and KD = 1.53 microM for HeLa/Fe5, were obtained from competition experiments. We conclude that specific binding sites for ferric citrate are constitutively expressed in plasma membrane and that their expres...Continue Reading

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Citations

Aug 15, 2000·Annual Review of Nutrition·M Wessling-Resnick
Mar 13, 2003·Redox Report : Communications in Free Radical Research·Jennifer D Ly, Alfons Lawen
May 5, 1999·Current Opinion in Chemical Biology·P AisenE A Leibold
Aug 15, 2006·Blood Cells, Molecules & Diseases·Jan KovarJiri Horak
Jun 1, 2005·Journal of Investigative Medicine : the Official Publication of the American Federation for Clinical Research·Oyebode OlakanmiBradley E Britigan
May 25, 1999·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·J M May
Jul 13, 2002·The Journal of Experimental Zoology·Amy G AslamkhanGregory A Ahearn
Jul 31, 1998·Analytical Biochemistry·L Brunner-DöpperH Goldenberg

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