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Fractionation of Tetrahymena ciliary membranes with triton X-114 and the identification of a ciliary membrane ATPase
Cilia were isolated from Tetrahymena thermophila, extracted with Triton X-114, and the detergent-soluble membrane + matrix proteins separated into Triton X-114 aqueous and detergent phases. The aqueous phase polypeptides include a high molecular mass polypeptide previously identified as a membrane d...
Formato: | Texto |
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Lenguaje: | English |
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The Rockefeller University Press
1988
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2115688/ https://www.ncbi.nlm.nih.gov/pubmed/2974460 |
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collection | PubMed |
description | Cilia were isolated from Tetrahymena thermophila, extracted with Triton X-114, and the detergent-soluble membrane + matrix proteins separated into Triton X-114 aqueous and detergent phases. The aqueous phase polypeptides include a high molecular mass polypeptide previously identified as a membrane dynein, detergent-soluble alpha and beta tubulins, and numerous polypeptides distinct from those found in axonemes. Integral membrane proteins partition into the detergent phase and include two major polypeptides of 58 and 50 kD, a 49-kD polypeptide, and 5 polypeptides in relatively minor amounts. The major detergent phase polypeptides are PAS-positive and are phosphorylated in vivo. A membrane-associated ATPase, distinct from the dynein-like protein, partitions into the Triton X-114 detergent phase and contains nearly 20% of the total ciliary ATPase activity. The ATPase requires Mg++ or Ca++ and is not inhibited by ouabain or vanadate. This procedure provides a gentle and rapid technique to separate integral membrane proteins from those that may be peripherally associated with the matrix or membrane. |
format | Text |
id | pubmed-2115688 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 1988 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-21156882008-05-01 Fractionation of Tetrahymena ciliary membranes with triton X-114 and the identification of a ciliary membrane ATPase J Cell Biol Articles Cilia were isolated from Tetrahymena thermophila, extracted with Triton X-114, and the detergent-soluble membrane + matrix proteins separated into Triton X-114 aqueous and detergent phases. The aqueous phase polypeptides include a high molecular mass polypeptide previously identified as a membrane dynein, detergent-soluble alpha and beta tubulins, and numerous polypeptides distinct from those found in axonemes. Integral membrane proteins partition into the detergent phase and include two major polypeptides of 58 and 50 kD, a 49-kD polypeptide, and 5 polypeptides in relatively minor amounts. The major detergent phase polypeptides are PAS-positive and are phosphorylated in vivo. A membrane-associated ATPase, distinct from the dynein-like protein, partitions into the Triton X-114 detergent phase and contains nearly 20% of the total ciliary ATPase activity. The ATPase requires Mg++ or Ca++ and is not inhibited by ouabain or vanadate. This procedure provides a gentle and rapid technique to separate integral membrane proteins from those that may be peripherally associated with the matrix or membrane. The Rockefeller University Press 1988-12-01 /pmc/articles/PMC2115688/ /pubmed/2974460 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 | Articles Fractionation of Tetrahymena ciliary membranes with triton X-114 and the identification of a ciliary membrane ATPase |
title | Fractionation of Tetrahymena ciliary membranes with triton X-114 and the identification of a ciliary membrane ATPase |
title_full | Fractionation of Tetrahymena ciliary membranes with triton X-114 and the identification of a ciliary membrane ATPase |
title_fullStr | Fractionation of Tetrahymena ciliary membranes with triton X-114 and the identification of a ciliary membrane ATPase |
title_full_unstemmed | Fractionation of Tetrahymena ciliary membranes with triton X-114 and the identification of a ciliary membrane ATPase |
title_short | Fractionation of Tetrahymena ciliary membranes with triton X-114 and the identification of a ciliary membrane ATPase |
title_sort | fractionation of tetrahymena ciliary membranes with triton x-114 and the identification of a ciliary membrane atpase |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2115688/ https://www.ncbi.nlm.nih.gov/pubmed/2974460 |