Depletion of intracellular potassium arrests coated pit formation and receptor-mediated endocytosis in fibroblasts

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

Research output: Contribution to journalArticle

321 Citations (Scopus)

Abstract

Depletion of intracellular potassium (K+) caused a marked reduction in the rate of endocytosis of receptor-bound low density lipoprotein (LDL) and epidermal growth factor (EGF) in human fibroblasts. K+ could be depleted slowly by a 3-hr incubation of cells in isotonic K+-free buffer. Rapid K+ depletion was induced by incubation of cells for 5 min with hypotonic medium, followed by transfer to isotonic K+-free buffer. Within 30 min of this treatment, cellular K+ levels fell by more than 60%. When the K+ level fell below a threshold of 40% of normal, the number of coated pits declined by 80% and the rate of endocytosis of 125I-LDL decreased by 70 to 95% despite normal to increased receptor binding. Similar results were obtained with 125I-epidermal growth factor. Addition of KCI to the culture medium up to 2 hr after K+ depletion restored cellular K+ levels and returned endocytosis of 125I-LDL promptly to normal. RbCl was as effective as KCl, but CsCl, LiCI, and (CH3)4NCI had no effect. Restoration by KCI was blocked by ouabain, indicating that uptake via the Na+/K+ ATPase was required. These data demonstrate that depletion of intracellular K+ reversibly arrests coated pit formation and receptor-mediated endocytosis in human fibroblasts.

Original languageEnglish (US)
Pages (from-to)273-285
Number of pages13
JournalCell
Volume33
Issue number1
DOIs
StatePublished - 1983

Fingerprint

Fibroblasts
Endocytosis
LDL Lipoproteins
Potassium
Epidermal Growth Factor
Buffers
Cells
Ouabain
Restoration
Culture Media
Adenosine Triphosphatases
LDL Receptors

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

Depletion of intracellular potassium arrests coated pit formation and receptor-mediated endocytosis in fibroblasts. / Larkin, Janet M.; Brown, Michael S.; Goldstein, Joseph L.; Anderson, Richard G W.

In: Cell, Vol. 33, No. 1, 1983, p. 273-285.

Research output: Contribution to journalArticle

@article{d2260e780dc34e728150101d0ab3a25b,
title = "Depletion of intracellular potassium arrests coated pit formation and receptor-mediated endocytosis in fibroblasts",
abstract = "Depletion of intracellular potassium (K+) caused a marked reduction in the rate of endocytosis of receptor-bound low density lipoprotein (LDL) and epidermal growth factor (EGF) in human fibroblasts. K+ could be depleted slowly by a 3-hr incubation of cells in isotonic K+-free buffer. Rapid K+ depletion was induced by incubation of cells for 5 min with hypotonic medium, followed by transfer to isotonic K+-free buffer. Within 30 min of this treatment, cellular K+ levels fell by more than 60{\%}. When the K+ level fell below a threshold of 40{\%} of normal, the number of coated pits declined by 80{\%} and the rate of endocytosis of 125I-LDL decreased by 70 to 95{\%} despite normal to increased receptor binding. Similar results were obtained with 125I-epidermal growth factor. Addition of KCI to the culture medium up to 2 hr after K+ depletion restored cellular K+ levels and returned endocytosis of 125I-LDL promptly to normal. RbCl was as effective as KCl, but CsCl, LiCI, and (CH3)4NCI had no effect. Restoration by KCI was blocked by ouabain, indicating that uptake via the Na+/K+ ATPase was required. These data demonstrate that depletion of intracellular K+ reversibly arrests coated pit formation and receptor-mediated endocytosis in human fibroblasts.",
author = "Larkin, {Janet M.} and Brown, {Michael S.} and Goldstein, {Joseph L.} and Anderson, {Richard G W}",
year = "1983",
doi = "10.1016/0092-8674(83)90356-2",
language = "English (US)",
volume = "33",
pages = "273--285",
journal = "Cell",
issn = "0092-8674",
publisher = "Cell Press",
number = "1",

}

TY - JOUR

T1 - Depletion of intracellular potassium arrests coated pit formation and receptor-mediated endocytosis in fibroblasts

AU - Larkin, Janet M.

AU - Brown, Michael S.

AU - Goldstein, Joseph L.

AU - Anderson, Richard G W

PY - 1983

Y1 - 1983

N2 - Depletion of intracellular potassium (K+) caused a marked reduction in the rate of endocytosis of receptor-bound low density lipoprotein (LDL) and epidermal growth factor (EGF) in human fibroblasts. K+ could be depleted slowly by a 3-hr incubation of cells in isotonic K+-free buffer. Rapid K+ depletion was induced by incubation of cells for 5 min with hypotonic medium, followed by transfer to isotonic K+-free buffer. Within 30 min of this treatment, cellular K+ levels fell by more than 60%. When the K+ level fell below a threshold of 40% of normal, the number of coated pits declined by 80% and the rate of endocytosis of 125I-LDL decreased by 70 to 95% despite normal to increased receptor binding. Similar results were obtained with 125I-epidermal growth factor. Addition of KCI to the culture medium up to 2 hr after K+ depletion restored cellular K+ levels and returned endocytosis of 125I-LDL promptly to normal. RbCl was as effective as KCl, but CsCl, LiCI, and (CH3)4NCI had no effect. Restoration by KCI was blocked by ouabain, indicating that uptake via the Na+/K+ ATPase was required. These data demonstrate that depletion of intracellular K+ reversibly arrests coated pit formation and receptor-mediated endocytosis in human fibroblasts.

AB - Depletion of intracellular potassium (K+) caused a marked reduction in the rate of endocytosis of receptor-bound low density lipoprotein (LDL) and epidermal growth factor (EGF) in human fibroblasts. K+ could be depleted slowly by a 3-hr incubation of cells in isotonic K+-free buffer. Rapid K+ depletion was induced by incubation of cells for 5 min with hypotonic medium, followed by transfer to isotonic K+-free buffer. Within 30 min of this treatment, cellular K+ levels fell by more than 60%. When the K+ level fell below a threshold of 40% of normal, the number of coated pits declined by 80% and the rate of endocytosis of 125I-LDL decreased by 70 to 95% despite normal to increased receptor binding. Similar results were obtained with 125I-epidermal growth factor. Addition of KCI to the culture medium up to 2 hr after K+ depletion restored cellular K+ levels and returned endocytosis of 125I-LDL promptly to normal. RbCl was as effective as KCl, but CsCl, LiCI, and (CH3)4NCI had no effect. Restoration by KCI was blocked by ouabain, indicating that uptake via the Na+/K+ ATPase was required. These data demonstrate that depletion of intracellular K+ reversibly arrests coated pit formation and receptor-mediated endocytosis in human fibroblasts.

UR - http://www.scopus.com/inward/record.url?scp=0020626609&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0020626609&partnerID=8YFLogxK

U2 - 10.1016/0092-8674(83)90356-2

DO - 10.1016/0092-8674(83)90356-2

M3 - Article

C2 - 6147196

AN - SCOPUS:0020626609

VL - 33

SP - 273

EP - 285

JO - Cell

JF - Cell

SN - 0092-8674

IS - 1

ER -