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Mechanism of Peptide-induced Mast Cell Degranulation : Translocation and Patch-Clamp Studies
Substance P and other polycationic peptides are thought to stimulate mast cell degranulation via direct activation of G proteins. We investigated the ability of extracellularly applied substance P to translocate into mast cells and the ability of intracellularly applied substance P to stimulate degr...
Autores principales: | , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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The Rockefeller University Press
1998
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2229441/ https://www.ncbi.nlm.nih.gov/pubmed/9806967 |
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author | Lorenz, Dorothea Wiesner, Burkhard Zipper, Josef Winkler, Anett Krause, Eberhard Beyermann, Michael Lindau, Manfred Bienert, Michael |
author_facet | Lorenz, Dorothea Wiesner, Burkhard Zipper, Josef Winkler, Anett Krause, Eberhard Beyermann, Michael Lindau, Manfred Bienert, Michael |
author_sort | Lorenz, Dorothea |
collection | PubMed |
description | Substance P and other polycationic peptides are thought to stimulate mast cell degranulation via direct activation of G proteins. We investigated the ability of extracellularly applied substance P to translocate into mast cells and the ability of intracellularly applied substance P to stimulate degranulation. In addition, we studied by reverse transcription–-PCR whether substance P-specific receptors are present in the mast cell membrane. To study translocation, a biologically active and enzymatically stable fluorescent analogue of substance P was synthesized. A rapid, substance P receptor- and energy-independent uptake of this peptide into pertussis toxin-treated and -untreated mast cells was demonstrated using confocal laser scanning microscopy. The peptide was shown to localize preferentially on or inside the mast cell granules using electron microscopic autoradiography with (125)I-labeled all-D substance P and (3)H-labeled substance P. Cell membrane capacitance measurements using the patch-clamp technique demonstrated that intracellularly applied substance P induced calcium transients and activated mast cell exocytosis with a time delay that depended on peptide concentration (delay of 100–500 s at concentrations of substance P from 50 to 5 μM). Degranulation in response to intracellularly applied substance P was inhibited by GDPβS and pertussis toxin, suggesting that substance P acts via G protein activation. These results support the recently proposed model of a receptor-independent mechanism of peptide-induced mast cell degranulation, which assumes a direct interaction of peptides with G protein α subunits subsequent to their translocation across the plasma membrane. |
format | Text |
id | pubmed-2229441 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 1998 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-22294412008-04-22 Mechanism of Peptide-induced Mast Cell Degranulation : Translocation and Patch-Clamp Studies Lorenz, Dorothea Wiesner, Burkhard Zipper, Josef Winkler, Anett Krause, Eberhard Beyermann, Michael Lindau, Manfred Bienert, Michael J Gen Physiol Article Substance P and other polycationic peptides are thought to stimulate mast cell degranulation via direct activation of G proteins. We investigated the ability of extracellularly applied substance P to translocate into mast cells and the ability of intracellularly applied substance P to stimulate degranulation. In addition, we studied by reverse transcription–-PCR whether substance P-specific receptors are present in the mast cell membrane. To study translocation, a biologically active and enzymatically stable fluorescent analogue of substance P was synthesized. A rapid, substance P receptor- and energy-independent uptake of this peptide into pertussis toxin-treated and -untreated mast cells was demonstrated using confocal laser scanning microscopy. The peptide was shown to localize preferentially on or inside the mast cell granules using electron microscopic autoradiography with (125)I-labeled all-D substance P and (3)H-labeled substance P. Cell membrane capacitance measurements using the patch-clamp technique demonstrated that intracellularly applied substance P induced calcium transients and activated mast cell exocytosis with a time delay that depended on peptide concentration (delay of 100–500 s at concentrations of substance P from 50 to 5 μM). Degranulation in response to intracellularly applied substance P was inhibited by GDPβS and pertussis toxin, suggesting that substance P acts via G protein activation. These results support the recently proposed model of a receptor-independent mechanism of peptide-induced mast cell degranulation, which assumes a direct interaction of peptides with G protein α subunits subsequent to their translocation across the plasma membrane. The Rockefeller University Press 1998-11-01 /pmc/articles/PMC2229441/ /pubmed/9806967 Text en 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 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). |
spellingShingle | Article Lorenz, Dorothea Wiesner, Burkhard Zipper, Josef Winkler, Anett Krause, Eberhard Beyermann, Michael Lindau, Manfred Bienert, Michael Mechanism of Peptide-induced Mast Cell Degranulation : Translocation and Patch-Clamp Studies |
title | Mechanism of Peptide-induced Mast Cell Degranulation : Translocation and Patch-Clamp Studies |
title_full | Mechanism of Peptide-induced Mast Cell Degranulation : Translocation and Patch-Clamp Studies |
title_fullStr | Mechanism of Peptide-induced Mast Cell Degranulation : Translocation and Patch-Clamp Studies |
title_full_unstemmed | Mechanism of Peptide-induced Mast Cell Degranulation : Translocation and Patch-Clamp Studies |
title_short | Mechanism of Peptide-induced Mast Cell Degranulation : Translocation and Patch-Clamp Studies |
title_sort | mechanism of peptide-induced mast cell degranulation : translocation and patch-clamp studies |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2229441/ https://www.ncbi.nlm.nih.gov/pubmed/9806967 |
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