ATP is required for receptor-mediated endocytosis in intact cells

Sandra L. Schmid, Laura L. Carter

Research output: Contribution to journalArticle

158 Citations (Scopus)

Abstract

We have demonstrated a requirement for cellular ATP in the receptor-mediated endocytosis of transferrin. This has been accomplished using a novel assay for endocytosis based on acquisition of resistance to the membrane impermeable reducing agent, glutathione (GSH). Diferric-transferrin was conjugated to biotin via a cleavable disulfide bond and iodinated. Internalization of 125I-biotin-S-S-transferrin (125I-BSST) was quantitated by adsorption to avidin-Sepharose after treatment of cells with GSH. Receptor-mediated endocytosis of 125I-BSST was severely inhibited in ATP-depleted cells. Similar results were obtained when ATP was depleted by incubation of cells either under a N2-atmosphere or in the presence of NaN3 and NaF. The latter treatment, alone, also resulted in a loss of surface transferrin receptors which could not be correlated to reductions in cellular ATP. In contrast to the acquisition of GSH resistance, the apparent internalization of 125I-BSST as assessed by inaccessibility to antitransferrin antibodies reached control levels in ATP-depleted cells. Our biochemical and morphological data suggested that, although ATP is required for receptor-mediated endocytosis, in ATP-depleted cells ligands can become efficiently sequestered into deeply invaginated pits that are inaccessible to large probes such as antibodies, but remain accessible to small molecules such as GSH.

Original languageEnglish (US)
Pages (from-to)2307-2318
Number of pages12
JournalJournal of Cell Biology
Volume111
Issue number6 PART 1
StatePublished - Dec 1990

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Endocytosis
Adenosine Triphosphate
Transferrin
Biotin
Purinergic P2 Receptors
Sodium Azide
Transferrin Receptors
Avidin
Antibodies
Reducing Agents
Atmosphere
Disulfides
Sepharose
Adsorption
Glutathione
Ligands
Membranes

ASJC Scopus subject areas

  • Cell Biology

Cite this

ATP is required for receptor-mediated endocytosis in intact cells. / Schmid, Sandra L.; Carter, Laura L.

In: Journal of Cell Biology, Vol. 111, No. 6 PART 1, 12.1990, p. 2307-2318.

Research output: Contribution to journalArticle

Schmid, Sandra L. ; Carter, Laura L. / ATP is required for receptor-mediated endocytosis in intact cells. In: Journal of Cell Biology. 1990 ; Vol. 111, No. 6 PART 1. pp. 2307-2318.
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