Visualizing the functional architecture of the endocytic machinery
Clathrin-mediated endocytosis is an essential process that forms vesicles from the plasma membrane. Although most of the protein components of the endocytic protein machinery have been thoroughly characterized, their organization at the endocytic site is poorly understood. We developed a fluorescenc...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2015-02-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/04535 |
| _version_ | 1829096872032075776 |
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| author | Andrea Picco Markus Mund Jonas Ries François Nédélec Marko Kaksonen |
| author_facet | Andrea Picco Markus Mund Jonas Ries François Nédélec Marko Kaksonen |
| author_sort | Andrea Picco |
| collection | DOAJ |
| description | Clathrin-mediated endocytosis is an essential process that forms vesicles from the plasma membrane. Although most of the protein components of the endocytic protein machinery have been thoroughly characterized, their organization at the endocytic site is poorly understood. We developed a fluorescence microscopy method to track the average positions of yeast endocytic proteins in relation to each other with a time precision below 1 s and with a spatial precision of ∼10 nm. With these data, integrated with shapes of endocytic membrane intermediates and with superresolution imaging, we could visualize the dynamic architecture of the endocytic machinery. We showed how different coat proteins are distributed within the coat structure and how the assembly dynamics of N-BAR proteins relate to membrane shape changes. Moreover, we found that the region of actin polymerization is located at the base of the endocytic invagination, with the growing ends of filaments pointing toward the plasma membrane. |
| first_indexed | 2024-04-11T09:03:24Z |
| format | Article |
| id | doaj.art-ffe352d4b3cb44199f62b29ec4c00fba |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-11T09:03:24Z |
| publishDate | 2015-02-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-ffe352d4b3cb44199f62b29ec4c00fba2022-12-22T04:32:42ZengeLife Sciences Publications LtdeLife2050-084X2015-02-01410.7554/eLife.04535Visualizing the functional architecture of the endocytic machineryAndrea Picco0https://orcid.org/0000-0003-2548-9183Markus Mund1https://orcid.org/0000-0001-6449-743XJonas Ries2https://orcid.org/0000-0002-6640-9250François Nédélec3https://orcid.org/0000-0002-8141-5288Marko Kaksonen4https://orcid.org/0000-0003-3645-7689Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, GermanyCell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, GermanyCell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, GermanyCell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, GermanyCell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, GermanyClathrin-mediated endocytosis is an essential process that forms vesicles from the plasma membrane. Although most of the protein components of the endocytic protein machinery have been thoroughly characterized, their organization at the endocytic site is poorly understood. We developed a fluorescence microscopy method to track the average positions of yeast endocytic proteins in relation to each other with a time precision below 1 s and with a spatial precision of ∼10 nm. With these data, integrated with shapes of endocytic membrane intermediates and with superresolution imaging, we could visualize the dynamic architecture of the endocytic machinery. We showed how different coat proteins are distributed within the coat structure and how the assembly dynamics of N-BAR proteins relate to membrane shape changes. Moreover, we found that the region of actin polymerization is located at the base of the endocytic invagination, with the growing ends of filaments pointing toward the plasma membrane.https://elifesciences.org/articles/04535endocytosisactinlive-cell imaging |
| spellingShingle | Andrea Picco Markus Mund Jonas Ries François Nédélec Marko Kaksonen Visualizing the functional architecture of the endocytic machinery eLife endocytosis actin live-cell imaging |
| title | Visualizing the functional architecture of the endocytic machinery |
| title_full | Visualizing the functional architecture of the endocytic machinery |
| title_fullStr | Visualizing the functional architecture of the endocytic machinery |
| title_full_unstemmed | Visualizing the functional architecture of the endocytic machinery |
| title_short | Visualizing the functional architecture of the endocytic machinery |
| title_sort | visualizing the functional architecture of the endocytic machinery |
| topic | endocytosis actin live-cell imaging |
| url | https://elifesciences.org/articles/04535 |
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