Co-migration and internalization of transferrin and its receptor on K562 cells.

Caroline Enns, J. W. Larrick, H. Suomalainen, J. Schroder, H. H. Sussman

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

The incorporation of iron into human cells involves the binding of diferric transferrin to a specific cell surface receptor. We studied the process of endocytosis in K562, a human erythroid cell line, by using tetramethylrhodamine isothiocyanate-labeled transferrin (TRITC-transferrin) and fluorescein isothiocyanate-labeled Fab fragments of goat antireceptor IgG preparation (FITC-Fab-antitransferrin receptor antibody). Because the antireceptor antibody and transferrin bind to different sites on the transferrin receptor molecule it was possible to simultaneously and independently follow ligand and receptor. At 4 degrees C, the binding of TRITC-transferrin or FITC-Fab antitransferrin receptor antibody exhibited diffuse membrane fluorescence. At 20 degrees C, the binding of TRITC-transferrin was followed by the rapid formation of aggregates. However, the FITC-Fab antitransferrin receptor did not show similar aggregation at 20 degrees C unless transferrin was present. In the presence of transferrin, the FITC-Fab antitransferrin receptor antibody formed aggregates at the same sites and within the same time period as TRITC transferrin, indicating co-migration. Although the diffuse surface staining of either label was removed by proteolysis, the larger aggregates were not susceptible to enzyme degradation, indicating that they were intracellular. The internal location of the aggregates was also demonstrated using permeabilized cells that had been preincubated with transferrin and fixed with 4% paraformaldehyde. These cells showed aggregated receptor in the interior of the cell when reacted with fluorescein-labeled antibody to the receptor. This indicated that the transferrin and the transferrin receptor co-internalize and migrate to the same structures within the cell.

Original languageEnglish (US)
Pages (from-to)579-585
Number of pages7
JournalJournal of Cell Biology
Volume97
Issue number2
StatePublished - Aug 1983
Externally publishedYes

Fingerprint

Transferrin Receptors
K562 Cells
Transferrin
Fluorescein-5-isothiocyanate
Antibodies
Fluorescein
Erythroid Cells
Immunoglobulin Fab Fragments
Cell Surface Receptors
Endocytosis
Goats
Proteolysis
Iron
Immunoglobulin G
Fluorescence
Staining and Labeling
Ligands
Cell Line
Membranes
tetramethylrhodamine isothiocyanate

ASJC Scopus subject areas

  • Cell Biology

Cite this

Enns, C., Larrick, J. W., Suomalainen, H., Schroder, J., & Sussman, H. H. (1983). Co-migration and internalization of transferrin and its receptor on K562 cells. Journal of Cell Biology, 97(2), 579-585.

Co-migration and internalization of transferrin and its receptor on K562 cells. / Enns, Caroline; Larrick, J. W.; Suomalainen, H.; Schroder, J.; Sussman, H. H.

In: Journal of Cell Biology, Vol. 97, No. 2, 08.1983, p. 579-585.

Research output: Contribution to journalArticle

Enns, C, Larrick, JW, Suomalainen, H, Schroder, J & Sussman, HH 1983, 'Co-migration and internalization of transferrin and its receptor on K562 cells.', Journal of Cell Biology, vol. 97, no. 2, pp. 579-585.
Enns C, Larrick JW, Suomalainen H, Schroder J, Sussman HH. Co-migration and internalization of transferrin and its receptor on K562 cells. Journal of Cell Biology. 1983 Aug;97(2):579-585.
Enns, Caroline ; Larrick, J. W. ; Suomalainen, H. ; Schroder, J. ; Sussman, H. H. / Co-migration and internalization of transferrin and its receptor on K562 cells. In: Journal of Cell Biology. 1983 ; Vol. 97, No. 2. pp. 579-585.
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abstract = "The incorporation of iron into human cells involves the binding of diferric transferrin to a specific cell surface receptor. We studied the process of endocytosis in K562, a human erythroid cell line, by using tetramethylrhodamine isothiocyanate-labeled transferrin (TRITC-transferrin) and fluorescein isothiocyanate-labeled Fab fragments of goat antireceptor IgG preparation (FITC-Fab-antitransferrin receptor antibody). Because the antireceptor antibody and transferrin bind to different sites on the transferrin receptor molecule it was possible to simultaneously and independently follow ligand and receptor. At 4 degrees C, the binding of TRITC-transferrin or FITC-Fab antitransferrin receptor antibody exhibited diffuse membrane fluorescence. At 20 degrees C, the binding of TRITC-transferrin was followed by the rapid formation of aggregates. However, the FITC-Fab antitransferrin receptor did not show similar aggregation at 20 degrees C unless transferrin was present. In the presence of transferrin, the FITC-Fab antitransferrin receptor antibody formed aggregates at the same sites and within the same time period as TRITC transferrin, indicating co-migration. Although the diffuse surface staining of either label was removed by proteolysis, the larger aggregates were not susceptible to enzyme degradation, indicating that they were intracellular. The internal location of the aggregates was also demonstrated using permeabilized cells that had been preincubated with transferrin and fixed with 4{\%} paraformaldehyde. These cells showed aggregated receptor in the interior of the cell when reacted with fluorescein-labeled antibody to the receptor. This indicated that the transferrin and the transferrin receptor co-internalize and migrate to the same structures within the cell.",
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