PMID: 11336795May 5, 2001Paper

The effects of wild-type and mutant HFE expression upon cellular iron uptake in transfected human embryonic kidney cells

Biochimica Et Biophysica Acta
G P Feeney, M Worwood

Abstract

In order to measure the effects of HFE (haemochromatosis) upon iron uptake, stable expression of wild-type and C282Y, H63D and S65C mutant HFE cDNA was established in HEK 293 cells. Control cells were transfected with empty vector. Expression of HFE mRNA and protein was detected in the cell lines transfected with HFE cDNA, but not in the control cell line. The ferritin concentration in wild-type cells cultured in 40 microM ferric ammonium citrate was 69% of that in control cells and 81% of that in C282Y cells. The ferritin concentration in H63D cells was intermediate between wild-type and C282Y and the ferritin concentration in S65C cells was similar to wild-type cells. Uptake of transferrin-iron in wild-type, C282Y and control cells was measured over 45 min. The Hill coefficients for transferrin-iron uptake were similar. The V(max) for transferrin-iron uptake in wild-type cells was 59.5% of control cells and 69.5% of C282Y cells. Estimates of K(m) were 232 nM for wild-type cells, 338 nM for C282Y cells and 570 nM for controls. Transferrin receptor levels were lowered, but not significantly, in the HFE transfected cells. The results show that HFE reduces transferrin-iron uptake, probably as an uncompetitive inhibitor.

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Citations

Nov 30, 2002·Lancet·Adrian Bomford
Apr 12, 2002·Journal of Hepatology·Daniel F WallaceJames S Dooley
Jul 23, 2003·Proceedings of the National Academy of Sciences of the United States of America·An-Sheng ZhangCaroline A Enns
Dec 16, 2003·Proceedings of the National Academy of Sciences of the United States of America·Shunji TomatsuWilliam S Sly
May 11, 2002·Leukemia & Lymphoma·M Tevfik DorakMark Worwood
Feb 19, 2005·Neurobiology of Aging·Sang Y Lee, James R Connor
Feb 24, 2015·Molecular Immunology·Alexandre ReubenRéjean Lapointe
Mar 19, 2005·Genetics in Medicine : Official Journal of the American College of Medical Genetics·M Tevfik DorakMark Worwood
Dec 30, 2006·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Sang Y LeeJames R Connor
Mar 27, 2004·The Journal of Biological Chemistry·Paige S Davies, Caroline A Enns
Apr 2, 2004·The Journal of Biological Chemistry·Anthony M Giannetti, Pamela J Björkman

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