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Cryo-electron tomography reveals the structural diversity of cardiac proteins in their cellular context
Cardiovascular diseases are a leading cause of death worldwide, but our understanding of the underlying mechanisms is limited, in part because of the complexity of the cellular machinery that controls the heart muscle contraction cycle. Cryogenic electron tomography (cryo-ET) provides a way to visua...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10634850/ https://www.ncbi.nlm.nih.gov/pubmed/37961228 http://dx.doi.org/10.1101/2023.10.26.564098 |
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author | Woldeyes, Rahel A. Nishiga, Masataka Vander Roest, Alison S. Engel, Leeya Giri, Prerna Montenegro, Gabrielle C. Wu, Andrew C. Dunn, Alexander R. Spudich, James A. Bernstein, Daniel Schmid, Michael F. Wu, Joseph C. Chiu, Wah |
author_facet | Woldeyes, Rahel A. Nishiga, Masataka Vander Roest, Alison S. Engel, Leeya Giri, Prerna Montenegro, Gabrielle C. Wu, Andrew C. Dunn, Alexander R. Spudich, James A. Bernstein, Daniel Schmid, Michael F. Wu, Joseph C. Chiu, Wah |
author_sort | Woldeyes, Rahel A. |
collection | PubMed |
description | Cardiovascular diseases are a leading cause of death worldwide, but our understanding of the underlying mechanisms is limited, in part because of the complexity of the cellular machinery that controls the heart muscle contraction cycle. Cryogenic electron tomography (cryo-ET) provides a way to visualize diverse cellular machinery while preserving contextual information like subcellular localization and transient complex formation, but this approach has not been widely applied to the study of heart muscle cells (cardiomyocytes). Here, we deploy a platform for studying cardiovascular disease by combining cryo-ET with human induced pluripotent stem cell–derived cardiomyocytes (hiPSC-CMs). After developing a cryo-ET workflow for visualizing macromolecules in hiPSC-CMs, we reconstructed sub-nanometer resolution structures of the human thin filament, a central component of the contractile machinery. We also visualized a previously unobserved organization of a regulatory complex that connects muscle contraction to calcium signaling (the troponin complex), highlighting the value of our approach for interrogating the structures of cardiac proteins in their cellular context. |
format | Online Article Text |
id | pubmed-10634850 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Cold Spring Harbor Laboratory |
record_format | MEDLINE/PubMed |
spelling | pubmed-106348502023-11-13 Cryo-electron tomography reveals the structural diversity of cardiac proteins in their cellular context Woldeyes, Rahel A. Nishiga, Masataka Vander Roest, Alison S. Engel, Leeya Giri, Prerna Montenegro, Gabrielle C. Wu, Andrew C. Dunn, Alexander R. Spudich, James A. Bernstein, Daniel Schmid, Michael F. Wu, Joseph C. Chiu, Wah bioRxiv Article Cardiovascular diseases are a leading cause of death worldwide, but our understanding of the underlying mechanisms is limited, in part because of the complexity of the cellular machinery that controls the heart muscle contraction cycle. Cryogenic electron tomography (cryo-ET) provides a way to visualize diverse cellular machinery while preserving contextual information like subcellular localization and transient complex formation, but this approach has not been widely applied to the study of heart muscle cells (cardiomyocytes). Here, we deploy a platform for studying cardiovascular disease by combining cryo-ET with human induced pluripotent stem cell–derived cardiomyocytes (hiPSC-CMs). After developing a cryo-ET workflow for visualizing macromolecules in hiPSC-CMs, we reconstructed sub-nanometer resolution structures of the human thin filament, a central component of the contractile machinery. We also visualized a previously unobserved organization of a regulatory complex that connects muscle contraction to calcium signaling (the troponin complex), highlighting the value of our approach for interrogating the structures of cardiac proteins in their cellular context. Cold Spring Harbor Laboratory 2023-10-26 /pmc/articles/PMC10634850/ /pubmed/37961228 http://dx.doi.org/10.1101/2023.10.26.564098 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, for noncommercial purposes only, and only so long as attribution is given to the creator. |
spellingShingle | Article Woldeyes, Rahel A. Nishiga, Masataka Vander Roest, Alison S. Engel, Leeya Giri, Prerna Montenegro, Gabrielle C. Wu, Andrew C. Dunn, Alexander R. Spudich, James A. Bernstein, Daniel Schmid, Michael F. Wu, Joseph C. Chiu, Wah Cryo-electron tomography reveals the structural diversity of cardiac proteins in their cellular context |
title | Cryo-electron tomography reveals the structural diversity of cardiac proteins in their cellular context |
title_full | Cryo-electron tomography reveals the structural diversity of cardiac proteins in their cellular context |
title_fullStr | Cryo-electron tomography reveals the structural diversity of cardiac proteins in their cellular context |
title_full_unstemmed | Cryo-electron tomography reveals the structural diversity of cardiac proteins in their cellular context |
title_short | Cryo-electron tomography reveals the structural diversity of cardiac proteins in their cellular context |
title_sort | cryo-electron tomography reveals the structural diversity of cardiac proteins in their cellular context |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10634850/ https://www.ncbi.nlm.nih.gov/pubmed/37961228 http://dx.doi.org/10.1101/2023.10.26.564098 |
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