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Using antibodies against Dictyostelium membranes to identify an actin- binding membrane protein

Polyclonal antibodies made against Dictyostelium discoideum membranes were used to block the interaction of those membranes with actin. As expected, actin interacted mostly with the internal surface of the membrane, demonstrated by the fact that whole cells could only absorb out a minor fraction of...

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Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 1987
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2114537/
https://www.ncbi.nlm.nih.gov/pubmed/3818790
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description Polyclonal antibodies made against Dictyostelium discoideum membranes were used to block the interaction of those membranes with actin. As expected, actin interacted mostly with the internal surface of the membrane, demonstrated by the fact that whole cells could only absorb out a minor fraction of the blocking antibody. The antibody was used to show that the membrane component(s) which interacted with actin were probably integral; they could be extracted with detergent but not with solutions designed to extract peripheral membrane proteins. To identify the responsible protein(s), Western transfers of membranes were cut into fractions which were tested for their ability to absorb out the blocking activity of the antibody. We observed a single peak at a molecular weight of approximately 20,000, and thus conclude that a 20,000-mol-wt protein is a major integral membrane actin-binding protein in Dictyostelium. This approach to the identification of proteins involved in actin-membrane interaction has allowed us to make the first identification of an actin-binding membrane protein which is based on its activity in native membranes.
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spelling pubmed-21145372008-05-01 Using antibodies against Dictyostelium membranes to identify an actin- binding membrane protein J Cell Biol Articles Polyclonal antibodies made against Dictyostelium discoideum membranes were used to block the interaction of those membranes with actin. As expected, actin interacted mostly with the internal surface of the membrane, demonstrated by the fact that whole cells could only absorb out a minor fraction of the blocking antibody. The antibody was used to show that the membrane component(s) which interacted with actin were probably integral; they could be extracted with detergent but not with solutions designed to extract peripheral membrane proteins. To identify the responsible protein(s), Western transfers of membranes were cut into fractions which were tested for their ability to absorb out the blocking activity of the antibody. We observed a single peak at a molecular weight of approximately 20,000, and thus conclude that a 20,000-mol-wt protein is a major integral membrane actin-binding protein in Dictyostelium. This approach to the identification of proteins involved in actin-membrane interaction has allowed us to make the first identification of an actin-binding membrane protein which is based on its activity in native membranes. The Rockefeller University Press 1987-03-01 /pmc/articles/PMC2114537/ /pubmed/3818790 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
Using antibodies against Dictyostelium membranes to identify an actin- binding membrane protein
title Using antibodies against Dictyostelium membranes to identify an actin- binding membrane protein
title_full Using antibodies against Dictyostelium membranes to identify an actin- binding membrane protein
title_fullStr Using antibodies against Dictyostelium membranes to identify an actin- binding membrane protein
title_full_unstemmed Using antibodies against Dictyostelium membranes to identify an actin- binding membrane protein
title_short Using antibodies against Dictyostelium membranes to identify an actin- binding membrane protein
title_sort using antibodies against dictyostelium membranes to identify an actin- binding membrane protein
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2114537/
https://www.ncbi.nlm.nih.gov/pubmed/3818790