Protein structure and interactions elucidated with in-cell NMR for different cell cycle phases and in 3D human tissue models
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.
Authors
- Eva Ištvánková (ORCID: https://orcid.org/0000-0002-2452-4883)
- Silvie Foldynová-Trantírková (ORCID: https://orcid.org/0000-0001-8863-083X)
- Bohumil Fafílek (ORCID: https://orcid.org/0000-0002-3869-7937)
- Gustavo Rico-Llanos (ORCID: https://orcid.org/0000-0001-5991-848X)
- Jan Ryneš (ORCID: https://orcid.org/0000-0003-2091-390X)
- Kristyna Kamarytova
- Pavel Krejčı́ (ORCID: https://orcid.org/0000-0003-0618-9134)
- Lucia Banci (ORCID: https://orcid.org/0000-0003-0562-5774)
- Libor Macůrek (ORCID: https://orcid.org/0000-0002-0987-1238)
- Tomáš Loja (ORCID: https://orcid.org/0000-0002-9007-266X)
- Lukáš Trantı́rek (ORCID: https://orcid.org/0000-0001-5948-4837)
- Enrico Luchinat (ORCID: https://orcid.org/0000-0003-4183-4311)
- Letizia Barbieri (ORCID: https://orcid.org/0000-0002-7457-490X)
- Michaela Krafčíková (ORCID: https://orcid.org/0000-0002-1210-4785)
Institutions
- Central European Institute of Technology (CZ)
- Czech Academy of Sciences (CZ)
- Masaryk University (CZ)
- Central European Institute of Technology – Masaryk University (CZ)
- Czech Academy of Sciences, Institute of Animal Physiology and Genetics (CZ)
- University Hospital Brno (CZ)
- St. Anne's University Hospital Brno (CZ)
- Czech Academy of Sciences, Institute of Molecular Genetics (CZ)
- Interuniversity Consortium for Magnetic Resonance (IT)
- University of Florence (IT)
Publication Details
- Journal
- Communications Biology
- Published
- 2025-02-07
- DOI
- https://doi.org/10.1038/s42003-025-07607-w
- Citations
- 8
- Primary Topic
- Advanced MRI Techniques and Applications
- Type
- article
- Field-Weighted Citation Impact
- 10.88
Funders
- European Commission
- Ministerstvo Školství, Mládeže a Tělovýchovy
- Grantová Agentura České Republiky
- Akademie Věd České Republiky
- Central European Institute of Technology
- Masarykova Univerzita
- Lékařská fakulta, Masarykova univerzita
- European Regional Development Fund