QUANTITATIVE STUDIES ON ENZYMES IN STRUCTURES IN STRIATED MUSCLES BY LABELED INHIBITOR METHODS : II. Confirmation of Radioautographic Measurement by Liquid-Scintillation Counting

Fragments of mouse diaphragm and sternomastoid muscles were incubated in diisopropyl-fluorophosphate (DFP)-(3)H in conditions known to saturate all the available DFP-sensitive reaction sites. After being extensively washed, the enzyme acetylcholinesterase (AChase) was specifically reactivated by tre...

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Detalles Bibliográficos
Autores principales: Rogers, A. W., Barnard, E. A.
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 1969
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2107826/
https://www.ncbi.nlm.nih.gov/pubmed/5768869
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author Rogers, A. W.
Barnard, E. A.
author_facet Rogers, A. W.
Barnard, E. A.
author_sort Rogers, A. W.
collection PubMed
description Fragments of mouse diaphragm and sternomastoid muscles were incubated in diisopropyl-fluorophosphate (DFP)-(3)H in conditions known to saturate all the available DFP-sensitive reaction sites. After being extensively washed, the enzyme acetylcholinesterase (AChase) was specifically reactivated by treatment with pyridine-2-aldoxime methiodide (2-PAM). The radioactive DP-groups released into solution by 2-PAM were measured by liquid scintillation counting, and related to the known number of motor endplates present. Considerable difficulty was encountered in reducing the excess, adsorbed radioactivity to acceptable levels: long washing routines, extraction with organic solvents, and removing excess muscle fiber by microdissection were necessary. Six experiments gave a mean value of 2.4 x 10(7)molecules AChase per sternomastoid endplate, in reasonable agreement with the previously reported measurements by radioautography.
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spelling pubmed-21078262008-05-01 QUANTITATIVE STUDIES ON ENZYMES IN STRUCTURES IN STRIATED MUSCLES BY LABELED INHIBITOR METHODS : II. Confirmation of Radioautographic Measurement by Liquid-Scintillation Counting Rogers, A. W. Barnard, E. A. J Cell Biol Article Fragments of mouse diaphragm and sternomastoid muscles were incubated in diisopropyl-fluorophosphate (DFP)-(3)H in conditions known to saturate all the available DFP-sensitive reaction sites. After being extensively washed, the enzyme acetylcholinesterase (AChase) was specifically reactivated by treatment with pyridine-2-aldoxime methiodide (2-PAM). The radioactive DP-groups released into solution by 2-PAM were measured by liquid scintillation counting, and related to the known number of motor endplates present. Considerable difficulty was encountered in reducing the excess, adsorbed radioactivity to acceptable levels: long washing routines, extraction with organic solvents, and removing excess muscle fiber by microdissection were necessary. Six experiments gave a mean value of 2.4 x 10(7)molecules AChase per sternomastoid endplate, in reasonable agreement with the previously reported measurements by radioautography. The Rockefeller University Press 1969-06-01 /pmc/articles/PMC2107826/ /pubmed/5768869 Text en Copyright © 1969 by The Rockefeller University Press. 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
Rogers, A. W.
Barnard, E. A.
QUANTITATIVE STUDIES ON ENZYMES IN STRUCTURES IN STRIATED MUSCLES BY LABELED INHIBITOR METHODS : II. Confirmation of Radioautographic Measurement by Liquid-Scintillation Counting
title QUANTITATIVE STUDIES ON ENZYMES IN STRUCTURES IN STRIATED MUSCLES BY LABELED INHIBITOR METHODS : II. Confirmation of Radioautographic Measurement by Liquid-Scintillation Counting
title_full QUANTITATIVE STUDIES ON ENZYMES IN STRUCTURES IN STRIATED MUSCLES BY LABELED INHIBITOR METHODS : II. Confirmation of Radioautographic Measurement by Liquid-Scintillation Counting
title_fullStr QUANTITATIVE STUDIES ON ENZYMES IN STRUCTURES IN STRIATED MUSCLES BY LABELED INHIBITOR METHODS : II. Confirmation of Radioautographic Measurement by Liquid-Scintillation Counting
title_full_unstemmed QUANTITATIVE STUDIES ON ENZYMES IN STRUCTURES IN STRIATED MUSCLES BY LABELED INHIBITOR METHODS : II. Confirmation of Radioautographic Measurement by Liquid-Scintillation Counting
title_short QUANTITATIVE STUDIES ON ENZYMES IN STRUCTURES IN STRIATED MUSCLES BY LABELED INHIBITOR METHODS : II. Confirmation of Radioautographic Measurement by Liquid-Scintillation Counting
title_sort quantitative studies on enzymes in structures in striated muscles by labeled inhibitor methods : ii. confirmation of radioautographic measurement by liquid-scintillation counting
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2107826/
https://www.ncbi.nlm.nih.gov/pubmed/5768869
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