Use of Nramp2-transfected Chinese hamster ovary cells and reticulocytes from mk/mk mice to study iron transport mechanisms.

Experimental Hematology
An-Sheng ZhangP Ponka

Abstract

We investigated mechanisms involved in iron (Fe) transport by DMT1 (endosomal Fe(II) exporter, encoded by the Nramp2 gene) using wild-type Chinese hamster ovary (CHO) cells and Nramp2-transfected CHO cells, as well as reticulocytes from normal and mk/mk mice that have a defect in DMT1. CHO cells and reticulocytes were incubated with 59Fe bound to various ligands. The radioiron was present in its Fe(II) or Fe(III) forms or bound to transferrin (Tf), and the internalized 59Fe measured under varying experimental conditions. Additionally, 125I-Tf interaction with reticulocytes was investigated and 59Fe incorporation into their heme was determined. Hyperexpression of DMT1 in CHO cells greatly increases their capacity to acquire ferrous iron. Although CHO-Nramp2 cells showed an increase in Fe(III) uptake as compared to CHO cells, they transported Fe(III) with much lower efficacy than Fe(II). In addition to their defect in Fe uptake, mk/mk reticulocytes also showed a decrease in Tf receptor levels. Given that CHO cells acquire iron from Fe(II)-ascorbate with much higher rates than from Fe(III)-Tf, Tf-receptor levels represent the rate-limiting step in their iron uptake. As Fe(III) transport by CHO-Nramp2 cells can be inhibited by the ...Continue Reading

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Citations

Aug 5, 2010·The Journal of Biological Chemistry·Ningning ZhaoMitchell D Knutson
Apr 15, 2011·Anemia·Eugenia Prus, Eitan Fibach
Nov 10, 2015·Toxicology·Archita Venugopal MenonJonghan Kim
Aug 25, 2011·Trends in Molecular Medicine·Pierre BrissotOlivier Loréal
Apr 17, 2010·Experimental Hematology·Shan Soe-LinPrem Ponka
Jun 23, 2009·Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases·Fernando López-SotoMireya de la Garza
Jan 31, 2012·Biochimica Et Biophysica Acta·Caiyong Chen, Barry H Paw
Aug 23, 2011·Biochimica Et Biophysica Acta·Alex D SheftelPrem Ponka
Apr 20, 2011·BMC Evolutionary Biology·João V NevesPedro N S Rodrigues
Sep 18, 2010·Journal of the American College of Cardiology·Pradeep GujjaYukitaka Shizukuda
Jul 26, 2008·American Journal of Physiology. Renal Physiology·Tania VeutheyMarta Elena Roque
Apr 19, 2021·Journal of Molecular Biology·Aaron T Bozzi, Rachelle Gaudet

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