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Microtubule dynamics and glutathione metabolism in phagocytizing human polymorphonuclear leukocytes
Glutathione oxidants such as tertiary butyl hydroperoxide were shown previously to prevent microtubule assembly and cause breakdown of preassembled cytoplasmic microtubules in human polymorphonuclear leukocytes. The objectives of the present study were to determine the temporal relationship between...
Autores principales: | , , , , |
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Formato: | Texto |
Lenguaje: | English |
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The Rockefeller University Press
1978
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2109992/ https://www.ncbi.nlm.nih.gov/pubmed/10605449 |
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author | Burchill, B. R. Oliver, J. M. Pearson, C. B. Leinbach, E. D. Berlin, R. D. |
author_facet | Burchill, B. R. Oliver, J. M. Pearson, C. B. Leinbach, E. D. Berlin, R. D. |
author_sort | Burchill, B. R. |
collection | PubMed |
description | Glutathione oxidants such as tertiary butyl hydroperoxide were shown previously to prevent microtubule assembly and cause breakdown of preassembled cytoplasmic microtubules in human polymorphonuclear leukocytes. The objectives of the present study were to determine the temporal relationship between the attachment and ingestion of phagocytic particles and the assembly of microtubules, and simultaneously to quantify the levels of reduced glutathione and products of its oxidation as potential physiological regulators of assembly. Polymorphonuclear leukocytes from human peripheral blood were induced to phagocytize opsonized zymosan at 30 degrees C. Microtubule assembly was assessed in the electron microscope by direct counts of microtubules in thin sections through centrioles. Acid extracts were assayed for reduced glutathione (GSH) and oxidized glutathione (GSSG), by the sensitive enzymatic procedure of Tietze. Washed protein pellets were assayed for free sulfhydryl groups and for mixed protein disulfides with glutathione (protein-SSG) after borohydride splitting of the disulfide bond. Resting cells have few assembled microtubules. Phagocytosis induces a cycle of rapid assembly followed by disassembly. Assembly is initiated by particle contact and is maximal by 3 min of phagocytosis. Disassembly after 5-9 min of phagocytosis is preceded by a slow rise in GSSG and coincides with a rapid rise in protein-SSG. Protein-SSG also increases under conditions in which butyl hydroperoxide inhibits the assembly of microtubules that normally follows binding of concanavalin A to leukocyte cell surface receptors. No evidence for direct involvement of GSH in the induction of assembly was obtained. The formation of protein-SSG, however, emerges as a possible regulatory mechanism for the inhibition of microtubule assembly and induction of their disassembly. |
format | Text |
id | pubmed-2109992 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 1978 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-21099922008-05-01 Microtubule dynamics and glutathione metabolism in phagocytizing human polymorphonuclear leukocytes Burchill, B. R. Oliver, J. M. Pearson, C. B. Leinbach, E. D. Berlin, R. D. J Cell Biol Journal Article Glutathione oxidants such as tertiary butyl hydroperoxide were shown previously to prevent microtubule assembly and cause breakdown of preassembled cytoplasmic microtubules in human polymorphonuclear leukocytes. The objectives of the present study were to determine the temporal relationship between the attachment and ingestion of phagocytic particles and the assembly of microtubules, and simultaneously to quantify the levels of reduced glutathione and products of its oxidation as potential physiological regulators of assembly. Polymorphonuclear leukocytes from human peripheral blood were induced to phagocytize opsonized zymosan at 30 degrees C. Microtubule assembly was assessed in the electron microscope by direct counts of microtubules in thin sections through centrioles. Acid extracts were assayed for reduced glutathione (GSH) and oxidized glutathione (GSSG), by the sensitive enzymatic procedure of Tietze. Washed protein pellets were assayed for free sulfhydryl groups and for mixed protein disulfides with glutathione (protein-SSG) after borohydride splitting of the disulfide bond. Resting cells have few assembled microtubules. Phagocytosis induces a cycle of rapid assembly followed by disassembly. Assembly is initiated by particle contact and is maximal by 3 min of phagocytosis. Disassembly after 5-9 min of phagocytosis is preceded by a slow rise in GSSG and coincides with a rapid rise in protein-SSG. Protein-SSG also increases under conditions in which butyl hydroperoxide inhibits the assembly of microtubules that normally follows binding of concanavalin A to leukocyte cell surface receptors. No evidence for direct involvement of GSH in the induction of assembly was obtained. The formation of protein-SSG, however, emerges as a possible regulatory mechanism for the inhibition of microtubule assembly and induction of their disassembly. The Rockefeller University Press 1978-02-01 /pmc/articles/PMC2109992/ /pubmed/10605449 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 | Journal Article Burchill, B. R. Oliver, J. M. Pearson, C. B. Leinbach, E. D. Berlin, R. D. Microtubule dynamics and glutathione metabolism in phagocytizing human polymorphonuclear leukocytes |
title | Microtubule dynamics and glutathione metabolism in phagocytizing human polymorphonuclear leukocytes |
title_full | Microtubule dynamics and glutathione metabolism in phagocytizing human polymorphonuclear leukocytes |
title_fullStr | Microtubule dynamics and glutathione metabolism in phagocytizing human polymorphonuclear leukocytes |
title_full_unstemmed | Microtubule dynamics and glutathione metabolism in phagocytizing human polymorphonuclear leukocytes |
title_short | Microtubule dynamics and glutathione metabolism in phagocytizing human polymorphonuclear leukocytes |
title_sort | microtubule dynamics and glutathione metabolism in phagocytizing human polymorphonuclear leukocytes |
topic | Journal Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2109992/ https://www.ncbi.nlm.nih.gov/pubmed/10605449 |
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