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J. Biol. Chem., Vol. 265, Issue 6, 3139-3145, Feb, 1990
A Sturrock, J Alexander, J Lamb, CM Craven and J Kaplan
HeLa cells incubated in serum-free medium accumulated 59Fe ("non-
transferrin iron") when incubated with either 59Fe-citrate, 59Fe-
nitrilotriacetate, or 59Fe dissolved in Tricine ascorbate. Accumulation of
iron was time-, concentration-, and Ca2+-dependent and was saturable.
Uptake of non-transferrin (non-Tf) iron was transferrin- independent
because of the fact that uptake occurred at pH 5.5, a pH at which
transferrin binds iron poorly and at which transferrin is not internalized
by cells. Uptake of non-Tf iron was less affected than uptake of
transferrin iron by 1) exposure of cells to trypsin, a maneuver that
cleaves Tf receptors, or 2) incubation of cells with phenylarsine oxide, an
agent that inhibits both fluid- and receptor- mediated internalization.
After exposure of cells to non-Tf iron at 37 degrees C, most of the
cell-associated radioactivity was recovered in heme and ferritin,
demonstrating that iron gained access to intracellular compartments and was
not simply adsorbed to the cell surface. Uptake of non-Tf iron could be
partially blocked by Cu2+ in a dose-dependent manner, while the
accumulation of transferrin-bound iron was unaffected by Cu2+. Other
transition metals, such as Zn2+, Cd2+, and Mn2+ were able to inhibit the
uptake of non-Tf iron to different degrees. The accumulation of 109Cd was
inhibited by incubation of cells with non-Tf iron, Cu2+, or Mn2+. The
extent of inhibition was concentration- and metal-dependent. A number of
cultured cell lines including HeLa, human skin fibroblasts, and Chinese
hamster ovary cells demonstrated uptake of non-Tf iron and 109Cd.
Additionally, an endosome acidification mutant of Chinese hamster ovary
cells, which exhibited an increase in non-Tf iron uptake, also exhibited an
increase in the uptake of Cd2+. These observations suggest that the
characteristics of the non-Tf iron transport system in HeLa cells are
similar if not identical to those reported for perfused rat liver (Wright,
T. L., Brissot, P., Ma, W.-L., and Weisiger, P. A. (1986) J. Biol. Chem.
261, 10909-10914) and suggest the existence of a family of transition metal
transport systems, each with a different metal specificity.
Characterization of a transferrin-independent uptake system for iron in HeLa cells
Department of Pathology, University of Utah School of Medicine, Salt Lake City 84132.
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