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Am J Physiol Cell Physiol (August 22, 2002). doi:10.1152/ajpcell.00184.2002
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Articles in PresS, published online ahead of print August 22, 2002
Am J Physiol Cell Physiol, 10.1152/ajpcell.00184.2002
Submitted on April 22, 2002
Accepted on August 19, 2002

EFFECT OF DMT1 KNOCKDOWN ON IRON, CADMIUM AND LEAD UPTAKE IN CACO-2 CELLS

Desmond I Bannon1, Roger Abounader2, Peter S J Lees3, and Joseph P Bressler4*

1 Environmental Health Sciences, Bloomberg School of Public Health, Baltimore, MD, USA; Department of Neurology, Kennedy Krieger Institute, Baltimore, MD, USA
2 Department of Neurology, Kennedy Krieger Institute, Baltimore, MD, USA; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD, USA
3 Environmental Health Sciences, Bloomberg School of Public Health, Baltimore, MD, USA
4 Department of Neurology, Kennedy Krieger Institute, Baltimore, MD, USA; Environmental Health Sciences, Bloomberg School of Public Health, Baltimore, MD, USA

* To whom correspondence should be addressed. E-mail: bressler{at}kennedykrieger.org.

DMT1 (divalent metal transporter), is a hydrogen-coupled divalent-metal transporter with a substrate preference for iron, although the protein when expressed in frog oocytes transports a broad range of metals, which includes the toxic metals cadmium and lead. Wild-type Caco-2 cells displayed saturable transport of lead and iron that was stimulated by acid. Cadmium and manganese inhibited transport of iron but zinc and lead did not. The involvement of DMT1 in the transport of toxic metals was examined by establishing clonal DMT1 knockdown and control Caco-2 cell lines. Knockdown cell lines displayed much lower levels of DMT1 mRNA and a smaller Vmax for iron uptake compared to control cell lines. One clone was further characterized and found to display an approximately 50% reduction in uptake of iron across a pH range from 5.5 to 7.4. Uptake for cadmium also decreased 50% across the same pH range but not for lead. These results show that DMT1 is important in iron and cadmium transport in Caco-2 cells, but that lead enters these cells through an independent hydrogen-driven mechanism.




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