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Am J Physiol Cell Physiol (January 2, 2002). doi:10.1152/ajpcell.00415.2001
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Articles in PresS, published online ahead of print January 2, 2002
Am J Physiol Cell Physiol, 10.1152/ajpcell.00415.2001
Submitted on August 27, 2001
Accepted on December 13, 2001

Differential regulation of transferrin-induced endocytosis by wild-type and C282Y-mutant HFE in transfected HeLa cells

Lukas Schwake1*, Andreas W Henkel2, Hans D Riedel1, Thorsten Schlenker1, Matthias Both2, Andrea Migala2, Boris Hadaschik1, Nataly Henfling1, and Wolfgang Stremmel1

1 Internal Medicine IV, Medical University Heidelberg, Heidelberg, Germany
2 Biomedical Optics, Max-Planck-Institute for Medical Research, Heidelberg, Germany

* To whom correspondence should be addressed. E-mail: base1971{at}hotmail.com.

The hereditary hemochromatosis protein HFE is known to complex with the transferrin receptor, however, its function regarding endocytosis of transferrin is unclear. We performed patch-clamp capacitance measurements in transfected HeLa cells carrying wild-type or C282Y-mutant HFE cDNA under the control of a tetracycline-sensitive promotor. Whole cell experiments in cells with suppressed expression of wild-type HFE revealed a decrease in membrane capacitance reflecting predominance of endocytosis in the presence of transferrin. Cells overexpressing C282Y-mutant HFE displayed less intense capacitance decreases, whereas no significant decrease was observed in cells overexpressing wild-type HFE. The formation of single endocytic vesicles in cells with suppressed expression of wild-type HFE was greatly increased in the presence of transferrin as revealed by cell-attached recordings. According to their calculated diameters, many of these vesicles corresponded to clathrin-coated vesicles. These results suggest that wild-type HFE negatively modulates the endocytic uptake of transferrin. This inhibitory effect is attenuated in cells expressing C282Y-mutant HFE. Time-resolved measurements of cell membrane capacitance provide a powerful tool to study transferrin-induced endocytosis in single cells.




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