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Comparative analysis of adaptor-mediated clathrin assembly reveals general principles for adaptor clustering
Clathrin-mediated endocytosis (CME) manages the sorting and uptake of the bulk of membrane proteins (or cargo) from the plasma membrane. CME is initiated by the formation of clathrin-coated pits (CCPs), in which adaptors nucleate clathrin assembly. Clathrin adaptors display diversity in both the typ...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5063622/ https://www.ncbi.nlm.nih.gov/pubmed/27559129 http://dx.doi.org/10.1091/mbc.E16-06-0399 |
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author | Pucadyil, Thomas J. Holkar, Sachin S. |
author_facet | Pucadyil, Thomas J. Holkar, Sachin S. |
author_sort | Pucadyil, Thomas J. |
collection | PubMed |
description | Clathrin-mediated endocytosis (CME) manages the sorting and uptake of the bulk of membrane proteins (or cargo) from the plasma membrane. CME is initiated by the formation of clathrin-coated pits (CCPs), in which adaptors nucleate clathrin assembly. Clathrin adaptors display diversity in both the type and number of evolutionarily conserved clathrin-binding boxes. How this diversity relates to the process of adaptor clustering as clathrin assembles around a growing pit remains unclear. Using real-time, fluorescence microscopy–based assays, we compare the formation kinetics and distribution of clathrin assemblies on membranes that display five unique clathrin adaptors. Correlations between equilibrium and kinetic parameters of clathrin assembly to the eventual adaptor distribution indicate that adaptor clustering is determined not by the amount of clathrin recruited or the degree of clathrin clustered but instead by the rate of clathrin assembly. Together our results emphasize the need to analyze kinetics of protein interactions to better understand mechanisms that regulate CME. |
format | Online Article Text |
id | pubmed-5063622 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-50636222016-12-30 Comparative analysis of adaptor-mediated clathrin assembly reveals general principles for adaptor clustering Pucadyil, Thomas J. Holkar, Sachin S. Mol Biol Cell Articles Clathrin-mediated endocytosis (CME) manages the sorting and uptake of the bulk of membrane proteins (or cargo) from the plasma membrane. CME is initiated by the formation of clathrin-coated pits (CCPs), in which adaptors nucleate clathrin assembly. Clathrin adaptors display diversity in both the type and number of evolutionarily conserved clathrin-binding boxes. How this diversity relates to the process of adaptor clustering as clathrin assembles around a growing pit remains unclear. Using real-time, fluorescence microscopy–based assays, we compare the formation kinetics and distribution of clathrin assemblies on membranes that display five unique clathrin adaptors. Correlations between equilibrium and kinetic parameters of clathrin assembly to the eventual adaptor distribution indicate that adaptor clustering is determined not by the amount of clathrin recruited or the degree of clathrin clustered but instead by the rate of clathrin assembly. Together our results emphasize the need to analyze kinetics of protein interactions to better understand mechanisms that regulate CME. The American Society for Cell Biology 2016-10-15 /pmc/articles/PMC5063622/ /pubmed/27559129 http://dx.doi.org/10.1091/mbc.E16-06-0399 Text en © 2016 Pucadyil and Holkar. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. |
spellingShingle | Articles Pucadyil, Thomas J. Holkar, Sachin S. Comparative analysis of adaptor-mediated clathrin assembly reveals general principles for adaptor clustering |
title | Comparative analysis of adaptor-mediated clathrin assembly reveals general principles for adaptor clustering |
title_full | Comparative analysis of adaptor-mediated clathrin assembly reveals general principles for adaptor clustering |
title_fullStr | Comparative analysis of adaptor-mediated clathrin assembly reveals general principles for adaptor clustering |
title_full_unstemmed | Comparative analysis of adaptor-mediated clathrin assembly reveals general principles for adaptor clustering |
title_short | Comparative analysis of adaptor-mediated clathrin assembly reveals general principles for adaptor clustering |
title_sort | comparative analysis of adaptor-mediated clathrin assembly reveals general principles for adaptor clustering |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5063622/ https://www.ncbi.nlm.nih.gov/pubmed/27559129 http://dx.doi.org/10.1091/mbc.E16-06-0399 |
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