Role of the coated endocytic vesicle in the uptake of receptor-bound low density lipoprotein in human fibroblasts

Richard G W Anderson, Michael S. Brown, Joseph L. Goldstein

Research output: Contribution to journalArticle

364 Citations (Scopus)

Abstract

125I-labeled and ferritin-labeled low density lipoprotein (LDL) were used as visual probes to study the surface distribution of LDL receptors and to examine the mechanism of the endocytosis of this lipoprotein in cultured human fibrobasts. Light microscopic autoradiograms of whole cells incubated with 125I-LDL at 4°C showed that LDL receptors were widely but unevenly distributed over the cell surface. With the electron microscope, we determined that 60-70% of the ferritin-labeled LDL that bound to cells at 4°C was localized over short coated segments of the plasma membrane that accounted for no more than 2% of the total surface area. To study the internalization process, cells were first allowed to bind ferritin-labeled LDL at 4°C and were then warmed to 37°C. Within 10 min, nearly all the surface-bound LDL-ferritin was incorporated into coated endocytic vesicles that were formed by the invagination and pinching-off of the coated membrane regions that contained the receptor-bound LDL. With increasing time at 37°C, these coated vesicles were observed sequentially to migrate through the cytoplasm (1 min), to lose their cytoplasmic coat (2 min), and to fuse with either primary or secondary lysosomes (6 min). The current data indicate that the coated regions of plasma membrane are specialized structures of rapid turnover that function to carry receptor-bound LDL, and perhaps other receptor-bound molecules, into the cell.

Original languageEnglish (US)
Pages (from-to)351-364
Number of pages14
JournalCell
Volume10
Issue number3
DOIs
StatePublished - 1977

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Coated Vesicles
Transport Vesicles
LDL Receptors
Fibroblasts
LDL Lipoproteins
Ferritins
Cell membranes
Cell Membrane
Endocytosis
Lysosomes
Lipoproteins
Cytoplasm
Electric fuses
Electrons
Light
Membranes
Electron microscopes
Molecules

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

Role of the coated endocytic vesicle in the uptake of receptor-bound low density lipoprotein in human fibroblasts. / Anderson, Richard G W; Brown, Michael S.; Goldstein, Joseph L.

In: Cell, Vol. 10, No. 3, 1977, p. 351-364.

Research output: Contribution to journalArticle

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AB - 125I-labeled and ferritin-labeled low density lipoprotein (LDL) were used as visual probes to study the surface distribution of LDL receptors and to examine the mechanism of the endocytosis of this lipoprotein in cultured human fibrobasts. Light microscopic autoradiograms of whole cells incubated with 125I-LDL at 4°C showed that LDL receptors were widely but unevenly distributed over the cell surface. With the electron microscope, we determined that 60-70% of the ferritin-labeled LDL that bound to cells at 4°C was localized over short coated segments of the plasma membrane that accounted for no more than 2% of the total surface area. To study the internalization process, cells were first allowed to bind ferritin-labeled LDL at 4°C and were then warmed to 37°C. Within 10 min, nearly all the surface-bound LDL-ferritin was incorporated into coated endocytic vesicles that were formed by the invagination and pinching-off of the coated membrane regions that contained the receptor-bound LDL. With increasing time at 37°C, these coated vesicles were observed sequentially to migrate through the cytoplasm (1 min), to lose their cytoplasmic coat (2 min), and to fuse with either primary or secondary lysosomes (6 min). The current data indicate that the coated regions of plasma membrane are specialized structures of rapid turnover that function to carry receptor-bound LDL, and perhaps other receptor-bound molecules, into the cell.

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