Protein structure and interactions elucidated with in-cell NMR for different cell cycle phases and in 3D human tissue models
Abstract Most of our knowledge of protein structure and function originates from experiments performed with purified proteins resuspended in dilute, buffered solutions. However, most proteins function in crowded intracellular environments with complex compositions. Significant efforts have been made...
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Nature Portfolio
2025-02-01
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| Series: | Communications Biology |
| Online Access: | https://doi.org/10.1038/s42003-025-07607-w |
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| author | Jan Rynes Eva Istvankova Michaela Dzurov Krafcikova Enrico Luchinat Letizia Barbieri Lucia Banci Kristyna Kamarytova Tomas Loja Bohumil Fafilek Gustavo Rico-Llanos Pavel Krejci Libor Macurek Silvie Foldynova-Trantirkova Lukas Trantirek |
| author_facet | Jan Rynes Eva Istvankova Michaela Dzurov Krafcikova Enrico Luchinat Letizia Barbieri Lucia Banci Kristyna Kamarytova Tomas Loja Bohumil Fafilek Gustavo Rico-Llanos Pavel Krejci Libor Macurek Silvie Foldynova-Trantirkova Lukas Trantirek |
| author_sort | Jan Rynes |
| collection | DOAJ |
| description | Abstract Most of our knowledge of protein structure and function originates from experiments performed with purified proteins resuspended in dilute, buffered solutions. However, most proteins function in crowded intracellular environments with complex compositions. Significant efforts have been made to develop tools to study proteins in their native cellular settings. Among these tools, in-cell NMR spectroscopy has been the sole technique for characterizing proteins in the intracellular space of living cells at atomic resolution and physiological temperature. Nevertheless, due to technological constraints, in-cell NMR studies have been limited to asynchronous single-cell suspensions, precluding obtaining information on protein behavior in different cellular states. In this study, we present a methodology that allows for obtaining an atomically resolved NMR readout of protein structure and interactions in living human cells synchronized in specific cell cycle phases and within 3D models of human tissue. The described approach opens avenues for investigating how protein structure or drug recognition responds to cell-cell communication or changes in intracellular space composition during transitions among cell cycle phases. |
| format | Article |
| id | doaj-art-0c7ab2d2f39b4ba082c83f341a0c2479 |
| institution | Kabale University |
| issn | 2399-3642 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Communications Biology |
| spelling | doaj-art-0c7ab2d2f39b4ba082c83f341a0c24792025-02-09T12:50:44ZengNature PortfolioCommunications Biology2399-36422025-02-01811910.1038/s42003-025-07607-wProtein structure and interactions elucidated with in-cell NMR for different cell cycle phases and in 3D human tissue modelsJan Rynes0Eva Istvankova1Michaela Dzurov Krafcikova2Enrico Luchinat3Letizia Barbieri4Lucia Banci5Kristyna Kamarytova6Tomas Loja7Bohumil Fafilek8Gustavo Rico-Llanos9Pavel Krejci10Libor Macurek11Silvie Foldynova-Trantirkova12Lukas Trantirek13Central European Institute of Technology, Masaryk UniversityCentral European Institute of Technology, Masaryk UniversityCentral European Institute of Technology, Masaryk UniversityInteruniversity Consortium for Magnetic Resonance of Metallo ProteinsInteruniversity Consortium for Magnetic Resonance of Metallo ProteinsInteruniversity Consortium for Magnetic Resonance of Metallo ProteinsCentral European Institute of Technology, Masaryk UniversityCentral European Institute of Technology, Masaryk UniversityDepartment of Biology, Faculty of Medicine, Masaryk UniversityDepartment of Biology, Faculty of Medicine, Masaryk UniversityDepartment of Biology, Faculty of Medicine, Masaryk UniversityInstitute of Molecular Genetics, Czech Academy of SciencesCentral European Institute of Technology, Masaryk UniversityCentral European Institute of Technology, Masaryk UniversityAbstract Most of our knowledge of protein structure and function originates from experiments performed with purified proteins resuspended in dilute, buffered solutions. However, most proteins function in crowded intracellular environments with complex compositions. Significant efforts have been made to develop tools to study proteins in their native cellular settings. Among these tools, in-cell NMR spectroscopy has been the sole technique for characterizing proteins in the intracellular space of living cells at atomic resolution and physiological temperature. Nevertheless, due to technological constraints, in-cell NMR studies have been limited to asynchronous single-cell suspensions, precluding obtaining information on protein behavior in different cellular states. In this study, we present a methodology that allows for obtaining an atomically resolved NMR readout of protein structure and interactions in living human cells synchronized in specific cell cycle phases and within 3D models of human tissue. The described approach opens avenues for investigating how protein structure or drug recognition responds to cell-cell communication or changes in intracellular space composition during transitions among cell cycle phases.https://doi.org/10.1038/s42003-025-07607-w |
| spellingShingle | Jan Rynes Eva Istvankova Michaela Dzurov Krafcikova Enrico Luchinat Letizia Barbieri Lucia Banci Kristyna Kamarytova Tomas Loja Bohumil Fafilek Gustavo Rico-Llanos Pavel Krejci Libor Macurek Silvie Foldynova-Trantirkova Lukas Trantirek Protein structure and interactions elucidated with in-cell NMR for different cell cycle phases and in 3D human tissue models Communications Biology |
| title | Protein structure and interactions elucidated with in-cell NMR for different cell cycle phases and in 3D human tissue models |
| title_full | Protein structure and interactions elucidated with in-cell NMR for different cell cycle phases and in 3D human tissue models |
| title_fullStr | Protein structure and interactions elucidated with in-cell NMR for different cell cycle phases and in 3D human tissue models |
| title_full_unstemmed | Protein structure and interactions elucidated with in-cell NMR for different cell cycle phases and in 3D human tissue models |
| title_short | Protein structure and interactions elucidated with in-cell NMR for different cell cycle phases and in 3D human tissue models |
| title_sort | protein structure and interactions elucidated with in cell nmr for different cell cycle phases and in 3d human tissue models |
| url | https://doi.org/10.1038/s42003-025-07607-w |
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