Receptor-mediated uptake of ferritin-bound iron by human intestinal Caco-2 cells

Swati Kalgaonkar, Bo Lönnerdal

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

Ferritin (Ft) is a large iron (Fe)-binding protein (∼450 kDa) that is found in plant and animal cells and can sequester up to 4500 Fe atoms per Ft molecule. Our previous studies on intestinal Caco-2 cells have shown that dietary factors affect the uptake of Fe from Ft in a manner different from that of Fe from FeSO4, suggesting a different mechanism for cellular uptake. The objective of this study was to determine the mechanism for Ft-Fe uptake using Caco-2 cells. Binding of 59Fe-labeled Ft at 4°C showed saturable kinetics, and Scatchard analysis resulted in a Kd of 1.6 μM, strongly indicating a receptor-mediated process. Competitive binding studies with excess unlabelled Ft significantly reduced binding, and uptake studies at 37°C showed saturation after 4 h. Enhancing and blocking endocytosis using Mas-7 (a G-protein activator) and hypertonic medium (0.5 M sucrose), respectively, demonstrated that Ft-Fe uptake by Mas-7-treated cells was 140% of control cells, whereas sucrose treatment resulted in a statistically significant reduction in Ft-Fe uptake by 70% as compared to controls. Inhibition of macropinocytosis with 5-(N,N-dimethyl)-amiloride (Na+/H+ antiport blocker) resulted in a decrease (by ∼20%) in Ft-Fe uptake at high concentrations of Ft, suggesting that enterocytes can use more than one Ft uptake mechanism in a concentration-dependent manner. These results suggest that Ft uptake by enterocytes is carried out via endocytosis when Ft levels are within a physiological range, whereas Ft at higher concentrations may be absorbed using the additional mechanism of macropinocytosis.

Original languageEnglish (US)
Pages (from-to)304-311
Number of pages8
JournalJournal of Nutritional Biochemistry
Volume20
Issue number4
DOIs
StatePublished - Apr 2009

Fingerprint

Caco-2 Cells
Ferritins
Iron
Enterocytes
Endocytosis
Sucrose
Iron-Binding Proteins
Cells
Competitive Binding
Ion Transport
Plant Cells
GTP-Binding Proteins

Keywords

  • Caco-2 cells
  • Ferritin
  • Ferritin receptor
  • Iron
  • Iron absorption

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry
  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism
  • Nutrition and Dietetics

Cite this

Receptor-mediated uptake of ferritin-bound iron by human intestinal Caco-2 cells. / Kalgaonkar, Swati; Lönnerdal, Bo.

In: Journal of Nutritional Biochemistry, Vol. 20, No. 4, 04.2009, p. 304-311.

Research output: Contribution to journalArticle

Kalgaonkar, Swati ; Lönnerdal, Bo. / Receptor-mediated uptake of ferritin-bound iron by human intestinal Caco-2 cells. In: Journal of Nutritional Biochemistry. 2009 ; Vol. 20, No. 4. pp. 304-311.
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