Nanoscale organization of the endogenous ASC speck
Summary: The NLRP3 inflammasome is a central component of the innate immune system. Its activation leads to formation of the ASC speck, a supramolecular assembly of the inflammasome adaptor protein ASC. Different models, based on ASC overexpression, have been proposed for the structure of the ASC sp...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-12-01
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| Series: | iScience |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2589004223024598 |
| _version_ | 1827582108554493952 |
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| author | Ivo M. Glück Grusha Primal Mathias Sebastian Strauss Virgile Rat Irene Gialdini Thomas Sebastian Ebert Che Stafford Ganesh Agam Suliana Manley Veit Hornung Ralf Jungmann Christian Sieben Don C. Lamb |
| author_facet | Ivo M. Glück Grusha Primal Mathias Sebastian Strauss Virgile Rat Irene Gialdini Thomas Sebastian Ebert Che Stafford Ganesh Agam Suliana Manley Veit Hornung Ralf Jungmann Christian Sieben Don C. Lamb |
| author_sort | Ivo M. Glück |
| collection | DOAJ |
| description | Summary: The NLRP3 inflammasome is a central component of the innate immune system. Its activation leads to formation of the ASC speck, a supramolecular assembly of the inflammasome adaptor protein ASC. Different models, based on ASC overexpression, have been proposed for the structure of the ASC speck. Using dual-color 3D super-resolution imaging (dSTORM and DNA-PAINT), we visualized the ASC speck structure following NLRP3 inflammasome activation using endogenous ASC expression. A complete structure was only obtainable by labeling with both anti-ASC antibodies and nanobodies. The complex varies in diameter between ∼800 and 1000 nm, and is composed of a dense core with emerging filaments. Dual-color confocal fluorescence microscopy indicated that the ASC speck does not colocalize with the microtubule-organizing center at late time points after Nigericin stimulation. From super-resolution images of whole cells, the ASC specks were sorted into a pseudo-time sequence indicating that they become denser but not larger during formation. |
| first_indexed | 2024-03-08T22:45:48Z |
| format | Article |
| id | doaj.art-f518dcc0b67d43c7ba16f7aa2b31c012 |
| institution | Directory Open Access Journal |
| issn | 2589-0042 |
| language | English |
| last_indexed | 2024-03-08T22:45:48Z |
| publishDate | 2023-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | iScience |
| spelling | doaj.art-f518dcc0b67d43c7ba16f7aa2b31c0122023-12-17T06:40:34ZengElsevieriScience2589-00422023-12-012612108382Nanoscale organization of the endogenous ASC speckIvo M. Glück0Grusha Primal Mathias1Sebastian Strauss2Virgile Rat3Irene Gialdini4Thomas Sebastian Ebert5Che Stafford6Ganesh Agam7Suliana Manley8Veit Hornung9Ralf Jungmann10Christian Sieben11Don C. Lamb12Department of Chemistry, Ludwig Maximilians-Universität München, Butenandtstraße 5-13, 81377 München, Germany; Center for Nano Science (CENS), Ludwig Maximilians-Universität München, Butenandtstraße 5-13, 81377 München, GermanyDepartment of Chemistry, Ludwig Maximilians-Universität München, Butenandtstraße 5-13, 81377 München, GermanyFaculty of Physics and Center for Nanoscience, Ludwig Maximilian University, Munich, Germany; Max Planck Institute of Biochemistry, Martinsried, GermanyDepartment of Chemistry, Ludwig Maximilians-Universität München, Butenandtstraße 5-13, 81377 München, Germany; Center for Nano Science (CENS), Ludwig Maximilians-Universität München, Butenandtstraße 5-13, 81377 München, GermanyDepartment of Chemistry, Ludwig Maximilians-Universität München, Butenandtstraße 5-13, 81377 München, Germany; Center for Nano Science (CENS), Ludwig Maximilians-Universität München, Butenandtstraße 5-13, 81377 München, GermanyGene Center and Department of Biochemistry, Ludwig-Maximilians-Universität, Munich, GermanyGene Center and Department of Biochemistry, Ludwig-Maximilians-Universität, Munich, GermanyDepartment of Chemistry, Ludwig Maximilians-Universität München, Butenandtstraße 5-13, 81377 München, Germany; Center for Nano Science (CENS), Ludwig Maximilians-Universität München, Butenandtstraße 5-13, 81377 München, GermanyLaboratory of Experimental Biophysics, École Polytechnique Fédérale de Lausanne, BSP 427 (Cubotron UNIL), Rte de la Sorge, CH-1015 Lausanne, SwitzerlandMax Planck Institute of Biochemistry, Martinsried, Germany; Gene Center and Department of Biochemistry, Ludwig-Maximilians-Universität, Munich, GermanyFaculty of Physics and Center for Nanoscience, Ludwig Maximilian University, Munich, Germany; Max Planck Institute of Biochemistry, Martinsried, GermanyLaboratory of Experimental Biophysics, École Polytechnique Fédérale de Lausanne, BSP 427 (Cubotron UNIL), Rte de la Sorge, CH-1015 Lausanne, Switzerland; Corresponding authorDepartment of Chemistry, Ludwig Maximilians-Universität München, Butenandtstraße 5-13, 81377 München, Germany; Center for Nano Science (CENS), Ludwig Maximilians-Universität München, Butenandtstraße 5-13, 81377 München, Germany; Corresponding authorSummary: The NLRP3 inflammasome is a central component of the innate immune system. Its activation leads to formation of the ASC speck, a supramolecular assembly of the inflammasome adaptor protein ASC. Different models, based on ASC overexpression, have been proposed for the structure of the ASC speck. Using dual-color 3D super-resolution imaging (dSTORM and DNA-PAINT), we visualized the ASC speck structure following NLRP3 inflammasome activation using endogenous ASC expression. A complete structure was only obtainable by labeling with both anti-ASC antibodies and nanobodies. The complex varies in diameter between ∼800 and 1000 nm, and is composed of a dense core with emerging filaments. Dual-color confocal fluorescence microscopy indicated that the ASC speck does not colocalize with the microtubule-organizing center at late time points after Nigericin stimulation. From super-resolution images of whole cells, the ASC specks were sorted into a pseudo-time sequence indicating that they become denser but not larger during formation.http://www.sciencedirect.com/science/article/pii/S2589004223024598ImmunologyBiotechnologyCell biology |
| spellingShingle | Ivo M. Glück Grusha Primal Mathias Sebastian Strauss Virgile Rat Irene Gialdini Thomas Sebastian Ebert Che Stafford Ganesh Agam Suliana Manley Veit Hornung Ralf Jungmann Christian Sieben Don C. Lamb Nanoscale organization of the endogenous ASC speck iScience Immunology Biotechnology Cell biology |
| title | Nanoscale organization of the endogenous ASC speck |
| title_full | Nanoscale organization of the endogenous ASC speck |
| title_fullStr | Nanoscale organization of the endogenous ASC speck |
| title_full_unstemmed | Nanoscale organization of the endogenous ASC speck |
| title_short | Nanoscale organization of the endogenous ASC speck |
| title_sort | nanoscale organization of the endogenous asc speck |
| topic | Immunology Biotechnology Cell biology |
| url | http://www.sciencedirect.com/science/article/pii/S2589004223024598 |
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