Cargando…

Massive endocytosis driven by lipidic forces originating in the outer plasmalemmal monolayer: a new approach to membrane recycling and lipid domains

The roles that lipids play in endocytosis are the subject of debate. Using electrical and imaging methods, we describe massive endocytosis (MEND) in baby hamster kidney (BHK) and HEK293 cells when the outer plasma membrane monolayer is perturbed by the nonionic detergents, Triton X-100 (TX100) and N...

Descripción completa

Detalles Bibliográficos
Autores principales: Fine, Michael, Llaguno, Marc C., Lariccia, Vincenzo, Lin, Mei-Jung, Yaradanakul, Alp, Hilgemann, Donald W.
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3032378/
https://www.ncbi.nlm.nih.gov/pubmed/21242300
http://dx.doi.org/10.1085/jgp.201010469
_version_ 1782197455671001088
author Fine, Michael
Llaguno, Marc C.
Lariccia, Vincenzo
Lin, Mei-Jung
Yaradanakul, Alp
Hilgemann, Donald W.
author_facet Fine, Michael
Llaguno, Marc C.
Lariccia, Vincenzo
Lin, Mei-Jung
Yaradanakul, Alp
Hilgemann, Donald W.
author_sort Fine, Michael
collection PubMed
description The roles that lipids play in endocytosis are the subject of debate. Using electrical and imaging methods, we describe massive endocytosis (MEND) in baby hamster kidney (BHK) and HEK293 cells when the outer plasma membrane monolayer is perturbed by the nonionic detergents, Triton X-100 (TX100) and NP-40. Some alkane detergents, the amphipathic drugs, edelfosine and tamoxifen, and the phospholipase inhibitor, U73122, are also effective. Uptake of the membrane tracer, FM 4–64, into vesicles and loss of reversible FM 4–64 binding confirm that 40–75% of the cell surface is internalized. Ongoing MEND stops in 2–4 s when amphipaths are removed, and amphipaths are without effect from the cytoplasmic side. Thus, expansion of the outer monolayer is critical. As found for Ca-activated MEND, vesicles formed are <100 nm in diameter, membrane ruffles are lost, and β-cyclodextrin treatments are inhibitory. However, amphipath-activated MEND does not require Ca transients, adenosine triphosphate (ATP) hydrolysis, G protein cycling, dynamins, or actin cytoskeleton remodeling. With elevated cytoplasmic ATP (>5 mM), MEND can reverse completely and be repeated multiple times in BHK and HEK293 cells, but not cardiac myocytes. Reversal is blocked by N-ethylmaleimide and a nitric oxide donor, nitroprusside. Constitutively expressed Na/Ca exchangers internalize roughly in proportion to surface membrane, whereas Na/K pump activities decrease over-proportionally. Sodium dodecyl sulfate and dodecylglucoside do not cause MEND during their application, but MEND occurs rapidly when they are removed. As monitored capacitively, the binding of these detergents decreases with MEND, whereas TX100 binding does not decrease. In summary, nonionic detergents can fractionate the plasma membrane in vivo, and vesicles formed connect immediately to physiological membrane-trafficking mechanisms. We suggest that lateral and transbilayer inhomogeneities of the plasma membrane provide potential energies that, when unbridled by triggers, can drive endocytosis by lipidic forces.
format Text
id pubmed-3032378
institution National Center for Biotechnology Information
language English
publishDate 2011
publisher The Rockefeller University Press
record_format MEDLINE/PubMed
spelling pubmed-30323782011-08-01 Massive endocytosis driven by lipidic forces originating in the outer plasmalemmal monolayer: a new approach to membrane recycling and lipid domains Fine, Michael Llaguno, Marc C. Lariccia, Vincenzo Lin, Mei-Jung Yaradanakul, Alp Hilgemann, Donald W. J Gen Physiol Article The roles that lipids play in endocytosis are the subject of debate. Using electrical and imaging methods, we describe massive endocytosis (MEND) in baby hamster kidney (BHK) and HEK293 cells when the outer plasma membrane monolayer is perturbed by the nonionic detergents, Triton X-100 (TX100) and NP-40. Some alkane detergents, the amphipathic drugs, edelfosine and tamoxifen, and the phospholipase inhibitor, U73122, are also effective. Uptake of the membrane tracer, FM 4–64, into vesicles and loss of reversible FM 4–64 binding confirm that 40–75% of the cell surface is internalized. Ongoing MEND stops in 2–4 s when amphipaths are removed, and amphipaths are without effect from the cytoplasmic side. Thus, expansion of the outer monolayer is critical. As found for Ca-activated MEND, vesicles formed are <100 nm in diameter, membrane ruffles are lost, and β-cyclodextrin treatments are inhibitory. However, amphipath-activated MEND does not require Ca transients, adenosine triphosphate (ATP) hydrolysis, G protein cycling, dynamins, or actin cytoskeleton remodeling. With elevated cytoplasmic ATP (>5 mM), MEND can reverse completely and be repeated multiple times in BHK and HEK293 cells, but not cardiac myocytes. Reversal is blocked by N-ethylmaleimide and a nitric oxide donor, nitroprusside. Constitutively expressed Na/Ca exchangers internalize roughly in proportion to surface membrane, whereas Na/K pump activities decrease over-proportionally. Sodium dodecyl sulfate and dodecylglucoside do not cause MEND during their application, but MEND occurs rapidly when they are removed. As monitored capacitively, the binding of these detergents decreases with MEND, whereas TX100 binding does not decrease. In summary, nonionic detergents can fractionate the plasma membrane in vivo, and vesicles formed connect immediately to physiological membrane-trafficking mechanisms. We suggest that lateral and transbilayer inhomogeneities of the plasma membrane provide potential energies that, when unbridled by triggers, can drive endocytosis by lipidic forces. The Rockefeller University Press 2011-02 /pmc/articles/PMC3032378/ /pubmed/21242300 http://dx.doi.org/10.1085/jgp.201010469 Text en © 2011 Fine et al. This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).
spellingShingle Article
Fine, Michael
Llaguno, Marc C.
Lariccia, Vincenzo
Lin, Mei-Jung
Yaradanakul, Alp
Hilgemann, Donald W.
Massive endocytosis driven by lipidic forces originating in the outer plasmalemmal monolayer: a new approach to membrane recycling and lipid domains
title Massive endocytosis driven by lipidic forces originating in the outer plasmalemmal monolayer: a new approach to membrane recycling and lipid domains
title_full Massive endocytosis driven by lipidic forces originating in the outer plasmalemmal monolayer: a new approach to membrane recycling and lipid domains
title_fullStr Massive endocytosis driven by lipidic forces originating in the outer plasmalemmal monolayer: a new approach to membrane recycling and lipid domains
title_full_unstemmed Massive endocytosis driven by lipidic forces originating in the outer plasmalemmal monolayer: a new approach to membrane recycling and lipid domains
title_short Massive endocytosis driven by lipidic forces originating in the outer plasmalemmal monolayer: a new approach to membrane recycling and lipid domains
title_sort massive endocytosis driven by lipidic forces originating in the outer plasmalemmal monolayer: a new approach to membrane recycling and lipid domains
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3032378/
https://www.ncbi.nlm.nih.gov/pubmed/21242300
http://dx.doi.org/10.1085/jgp.201010469
work_keys_str_mv AT finemichael massiveendocytosisdrivenbylipidicforcesoriginatingintheouterplasmalemmalmonolayeranewapproachtomembranerecyclingandlipiddomains
AT llagunomarcc massiveendocytosisdrivenbylipidicforcesoriginatingintheouterplasmalemmalmonolayeranewapproachtomembranerecyclingandlipiddomains
AT laricciavincenzo massiveendocytosisdrivenbylipidicforcesoriginatingintheouterplasmalemmalmonolayeranewapproachtomembranerecyclingandlipiddomains
AT linmeijung massiveendocytosisdrivenbylipidicforcesoriginatingintheouterplasmalemmalmonolayeranewapproachtomembranerecyclingandlipiddomains
AT yaradanakulalp massiveendocytosisdrivenbylipidicforcesoriginatingintheouterplasmalemmalmonolayeranewapproachtomembranerecyclingandlipiddomains
AT hilgemanndonaldw massiveendocytosisdrivenbylipidicforcesoriginatingintheouterplasmalemmalmonolayeranewapproachtomembranerecyclingandlipiddomains