Co-localization of 125I-epidermal growth factor and ferritin-low density lipoprotein in coated pits

A quantitative electron microscopic study in normal mutant human fibroblasts

Jean Louis Carpentier, Phillip Gorden, Richard G.W. Anderson, Joseph L. Goldstein, Michael S. Brown, Stanley Cohen, Lelio Orci

Research output: Contribution to journalArticle

112 Citations (Scopus)

Abstract

LOW density lipoprotein (LDL) and epidermal growth factor (EGF) bind to receptors on the surface of human fibroblasts and are internalized in coated vesicles. Each of the ligands has been studied separately by electron microscopy in human fibroblasts using ferritin-LDL as one visual probe and 125I-EGF as a second visual probe. A mutant strain of human fibroblasts (J.D.) has been described in which LDL does not localize to coated pits and hence is not internalized. Because LDL and EGF do not compete with each other for binding, in the current studies we coincubated the two ligands with normal and mutant cells to visualize their cellular fates. In normal fibroblasts ferritin-LDL and 125I-EGF both bound preferentially to coated pits at 4°C and both ligands were internalized into endocytotic vesicles and lysosomes. Quantitative studies in normal cells showed that 75% of the coated pits and vesicles that contained 125I-EGF also contained ferritin-LDL, indicating that both ligands enter the cell through the same endocytotic vesicles. In the LDL internalization-mutant J.D. cells, ferritin-LDL did not localize in coated pits and was not internalized, but 125I-EGF bound to coated pits and was internalized just as in normal fibroblasts.

Original languageEnglish (US)
Pages (from-to)73-77
Number of pages5
JournalJournal of Cell Biology
Volume95
Issue number1
DOIs
StatePublished - Oct 1 1982

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Ferritins
LDL Lipoproteins
Epidermal Growth Factor
Lipoproteins
Fibroblasts
Electrons
Coated Vesicles
Ligands
Lysosomes
Electron Microscopy

ASJC Scopus subject areas

  • Cell Biology

Cite this

Co-localization of 125I-epidermal growth factor and ferritin-low density lipoprotein in coated pits : A quantitative electron microscopic study in normal mutant human fibroblasts. / Carpentier, Jean Louis; Gorden, Phillip; Anderson, Richard G.W.; Goldstein, Joseph L.; Brown, Michael S.; Cohen, Stanley; Orci, Lelio.

In: Journal of Cell Biology, Vol. 95, No. 1, 01.10.1982, p. 73-77.

Research output: Contribution to journalArticle

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abstract = "LOW density lipoprotein (LDL) and epidermal growth factor (EGF) bind to receptors on the surface of human fibroblasts and are internalized in coated vesicles. Each of the ligands has been studied separately by electron microscopy in human fibroblasts using ferritin-LDL as one visual probe and 125I-EGF as a second visual probe. A mutant strain of human fibroblasts (J.D.) has been described in which LDL does not localize to coated pits and hence is not internalized. Because LDL and EGF do not compete with each other for binding, in the current studies we coincubated the two ligands with normal and mutant cells to visualize their cellular fates. In normal fibroblasts ferritin-LDL and 125I-EGF both bound preferentially to coated pits at 4°C and both ligands were internalized into endocytotic vesicles and lysosomes. Quantitative studies in normal cells showed that 75{\%} of the coated pits and vesicles that contained 125I-EGF also contained ferritin-LDL, indicating that both ligands enter the cell through the same endocytotic vesicles. In the LDL internalization-mutant J.D. cells, ferritin-LDL did not localize in coated pits and was not internalized, but 125I-EGF bound to coated pits and was internalized just as in normal fibroblasts.",
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