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PDZ interactions regulate rapid turnover of the scaffolding protein EBP50 in microvilli
Scaffolding proteins containing PDZ (postsynaptic density 95/discs large/zonula occludens-1) domains are believed to provide relatively stable linkages between components of macromolecular complexes and in some cases to bridge to the actin cytoskeleton. The microvillar scaffolding protein EBP50 (ERM...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Rockefeller University Press
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3410424/ https://www.ncbi.nlm.nih.gov/pubmed/22801783 http://dx.doi.org/10.1083/jcb.201204008 |
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author | Garbett, Damien Bretscher, Anthony |
author_facet | Garbett, Damien Bretscher, Anthony |
author_sort | Garbett, Damien |
collection | PubMed |
description | Scaffolding proteins containing PDZ (postsynaptic density 95/discs large/zonula occludens-1) domains are believed to provide relatively stable linkages between components of macromolecular complexes and in some cases to bridge to the actin cytoskeleton. The microvillar scaffolding protein EBP50 (ERM-binding phosphoprotein of 50 kD), consisting of two PDZ domains and an ezrin-binding site, retains specific proteins in microvilli and is necessary for microvillar biogenesis. Our analysis of the dynamics of microvillar proteins in vivo indicated that ezrin and microvillar membrane proteins had dynamics consistent with actin treadmilling and microvillar lifetimes. However, EBP50 was highly dynamic, turning over within seconds. EBP50 turnover was reduced by mutations that inactivate its PDZ domains and was enhanced by protein kinase C phosphorylation. Using a novel in vitro photoactivation fluorescence assay, the EBP50–ezrin interaction was shown to have a slow off-rate that was dramatically enhanced in a PDZ-regulated manner by addition of cell extract to near in vivo levels. Thus, the linking of relatively stable microvillar components can be mediated by surprisingly dynamic EBP50, a finding that may have important ramifications for other scaffolding proteins. |
format | Online Article Text |
id | pubmed-3410424 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-34104242013-01-23 PDZ interactions regulate rapid turnover of the scaffolding protein EBP50 in microvilli Garbett, Damien Bretscher, Anthony J Cell Biol Research Articles Scaffolding proteins containing PDZ (postsynaptic density 95/discs large/zonula occludens-1) domains are believed to provide relatively stable linkages between components of macromolecular complexes and in some cases to bridge to the actin cytoskeleton. The microvillar scaffolding protein EBP50 (ERM-binding phosphoprotein of 50 kD), consisting of two PDZ domains and an ezrin-binding site, retains specific proteins in microvilli and is necessary for microvillar biogenesis. Our analysis of the dynamics of microvillar proteins in vivo indicated that ezrin and microvillar membrane proteins had dynamics consistent with actin treadmilling and microvillar lifetimes. However, EBP50 was highly dynamic, turning over within seconds. EBP50 turnover was reduced by mutations that inactivate its PDZ domains and was enhanced by protein kinase C phosphorylation. Using a novel in vitro photoactivation fluorescence assay, the EBP50–ezrin interaction was shown to have a slow off-rate that was dramatically enhanced in a PDZ-regulated manner by addition of cell extract to near in vivo levels. Thus, the linking of relatively stable microvillar components can be mediated by surprisingly dynamic EBP50, a finding that may have important ramifications for other scaffolding proteins. The Rockefeller University Press 2012-07-23 /pmc/articles/PMC3410424/ /pubmed/22801783 http://dx.doi.org/10.1083/jcb.201204008 Text en © 2012 Garbett and Bretscher 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 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/). |
spellingShingle | Research Articles Garbett, Damien Bretscher, Anthony PDZ interactions regulate rapid turnover of the scaffolding protein EBP50 in microvilli |
title | PDZ interactions regulate rapid turnover of the scaffolding protein EBP50 in microvilli |
title_full | PDZ interactions regulate rapid turnover of the scaffolding protein EBP50 in microvilli |
title_fullStr | PDZ interactions regulate rapid turnover of the scaffolding protein EBP50 in microvilli |
title_full_unstemmed | PDZ interactions regulate rapid turnover of the scaffolding protein EBP50 in microvilli |
title_short | PDZ interactions regulate rapid turnover of the scaffolding protein EBP50 in microvilli |
title_sort | pdz interactions regulate rapid turnover of the scaffolding protein ebp50 in microvilli |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3410424/ https://www.ncbi.nlm.nih.gov/pubmed/22801783 http://dx.doi.org/10.1083/jcb.201204008 |
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