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Regulated motion of glycoproteins revealed by direct visualization of a single cargo in the endoplasmic reticulum
The quality of cargo proteins in the endoplasmic reticulum (ER) is affected by their motion during folding. To understand how the diffusion of secretory cargo proteins is regulated in the ER, we directly analyze the motion of a single cargo molecule using fluorescence imaging/fluctuation analyses. W...
Autores principales: | , , , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
The Rockefeller University Press
2008
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2213621/ https://www.ncbi.nlm.nih.gov/pubmed/18195104 http://dx.doi.org/10.1083/jcb.200704078 |
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author | Nagaya, Hisao Tamura, Taku Higa-Nishiyama, Arisa Ohashi, Koji Takeuchi, Mayumi Hashimoto, Hitoshi Hatsuzawa, Kiyotaka Kinjo, Masataka Okada, Tatsuya Wada, Ikuo |
author_facet | Nagaya, Hisao Tamura, Taku Higa-Nishiyama, Arisa Ohashi, Koji Takeuchi, Mayumi Hashimoto, Hitoshi Hatsuzawa, Kiyotaka Kinjo, Masataka Okada, Tatsuya Wada, Ikuo |
author_sort | Nagaya, Hisao |
collection | PubMed |
description | The quality of cargo proteins in the endoplasmic reticulum (ER) is affected by their motion during folding. To understand how the diffusion of secretory cargo proteins is regulated in the ER, we directly analyze the motion of a single cargo molecule using fluorescence imaging/fluctuation analyses. We find that the addition of two N-glycans onto the cargo dramatically alters their diffusion by transient binding to membrane components that are confined by hyperosmolarity. Via simultaneous observation of a single cargo and ER exit sites (ERESs), we could exclude ERESs as the binding sites. Remarkably, actin cytoskeleton was required for the transient binding. These results provide a molecular basis for hypertonicity-induced immobilization of cargo, which is dependent on glycosylation at multiple sites but not the completion of proper folding. We propose that diffusion of secretory glycoproteins in the ER lumen is controlled from the cytoplasm to reduce the chances of aggregation. |
format | Text |
id | pubmed-2213621 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-22136212008-07-14 Regulated motion of glycoproteins revealed by direct visualization of a single cargo in the endoplasmic reticulum Nagaya, Hisao Tamura, Taku Higa-Nishiyama, Arisa Ohashi, Koji Takeuchi, Mayumi Hashimoto, Hitoshi Hatsuzawa, Kiyotaka Kinjo, Masataka Okada, Tatsuya Wada, Ikuo J Cell Biol Research Articles The quality of cargo proteins in the endoplasmic reticulum (ER) is affected by their motion during folding. To understand how the diffusion of secretory cargo proteins is regulated in the ER, we directly analyze the motion of a single cargo molecule using fluorescence imaging/fluctuation analyses. We find that the addition of two N-glycans onto the cargo dramatically alters their diffusion by transient binding to membrane components that are confined by hyperosmolarity. Via simultaneous observation of a single cargo and ER exit sites (ERESs), we could exclude ERESs as the binding sites. Remarkably, actin cytoskeleton was required for the transient binding. These results provide a molecular basis for hypertonicity-induced immobilization of cargo, which is dependent on glycosylation at multiple sites but not the completion of proper folding. We propose that diffusion of secretory glycoproteins in the ER lumen is controlled from the cytoplasm to reduce the chances of aggregation. The Rockefeller University Press 2008-01-14 /pmc/articles/PMC2213621/ /pubmed/18195104 http://dx.doi.org/10.1083/jcb.200704078 Text en Copyright © 2008, 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 | Research Articles Nagaya, Hisao Tamura, Taku Higa-Nishiyama, Arisa Ohashi, Koji Takeuchi, Mayumi Hashimoto, Hitoshi Hatsuzawa, Kiyotaka Kinjo, Masataka Okada, Tatsuya Wada, Ikuo Regulated motion of glycoproteins revealed by direct visualization of a single cargo in the endoplasmic reticulum |
title | Regulated motion of glycoproteins revealed by direct visualization of a single cargo in the endoplasmic reticulum |
title_full | Regulated motion of glycoproteins revealed by direct visualization of a single cargo in the endoplasmic reticulum |
title_fullStr | Regulated motion of glycoproteins revealed by direct visualization of a single cargo in the endoplasmic reticulum |
title_full_unstemmed | Regulated motion of glycoproteins revealed by direct visualization of a single cargo in the endoplasmic reticulum |
title_short | Regulated motion of glycoproteins revealed by direct visualization of a single cargo in the endoplasmic reticulum |
title_sort | regulated motion of glycoproteins revealed by direct visualization of a single cargo in the endoplasmic reticulum |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2213621/ https://www.ncbi.nlm.nih.gov/pubmed/18195104 http://dx.doi.org/10.1083/jcb.200704078 |
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