Physics-based evolution of transmembrane helices reveals mechanisms of cholesterol attraction
The existence of linear cholesterol-recognition motifs in transmembrane domains has long been debated. Evolutionary molecular dynamics (Evo-MD) simulations-genetic algorithms guided by (coarse-grained) molecular force-fields-reveal that thermodynamic optimal cholesterol attraction in isolated alpha-helical transmembrane domains occurs when multiple consecutive lysine/arginine residues flank a short hydrophobic segment. These findings are supported by atomistic simulations and solid-state NMR experiments. Our analyses illustrate that linear motifs in transmembrane domains exhibit weak binding affinity for cholesterol, characterized by sub-microsecond residence times, challenging the predictive value of linear CRAC/CARC motifs for cholesterol binding. Membrane protein database analyses suggest even weaker affinity for native linear motifs, whereas live cell assays demonstrate that optimizing cholesterol binding restricts transmembrane domains to the endoplasmic reticulum post-translationally. In summary, these findings contribute to our understanding of cholesterol-protein interactions and offer insight into the mechanisms of protein-mediated cholesterol regulation within membranes.
Authors
- Roberto Sansevrino (ORCID: https://orcid.org/0009-0001-4464-5164)
- Loren B. Andreas (ORCID: https://orcid.org/0000-0003-3216-9065)
- Jeroen Methorst (ORCID: https://orcid.org/0000-0003-4507-4882)
- Partha Pyne (ORCID: https://orcid.org/0000-0002-9352-4510)
- Dragomir Milovanović (ORCID: https://orcid.org/0000-0002-6625-1879)
- Dennis Aschmann (ORCID: https://orcid.org/0009-0005-8889-0515)
- Nino Verwei (ORCID: https://orcid.org/0000-0003-2464-6896)
- Paweł Chodnicki (ORCID: https://orcid.org/0000-0003-4563-2469)
- Christian Hoffmann (ORCID: https://orcid.org/0000-0003-3478-2410)
- Jacek Czub (ORCID: https://orcid.org/0000-0003-3639-6935)
- Herre Jelger Risselada (ORCID: https://orcid.org/0000-0003-1410-6570)
- Niek van Hilten (ORCID: https://orcid.org/0000-0003-1204-2489)
- Alexander Kros (ORCID: https://orcid.org/0000-0002-3983-3048)
- H. Wang (ORCID: https://orcid.org/0009-0000-5517-3038)
Institutions
- Leiden University (NL)
- Gdańsk University of Technology (PL)
- University of California, San Francisco (US)
- TU Dortmund University (DE)
- University of Applied Sciences Leiden (NL)
- German Center for Neurodegenerative Diseases (DE)
- Humboldt-Universität zu Berlin (DE)
- University of Gdańsk (PL)
- Berlin Institute of Health at Charité - Universitätsmedizin Berlin (DE)
- Max Planck Institute for Multidisciplinary Sciences (DE)
Publication Details
- Journal
- Nature Communications
- Published
- 2025-10-20
- DOI
- https://doi.org/10.1038/s41467-025-63769-5
- Citations
- 2
- Primary Topic
- Lipid Membrane Structure and Behavior
- Type
- article
- Field-Weighted Citation Impact
- 1.61
Funders
- Deutsche Forschungsgemeinschaft
- Nederlandse Organisatie voor Wetenschappelijk Onderzoek
- Narodowym Centrum Nauki
- Deutsches Zentrum für Neurodegenerative Erkrankungen
- Infrastruktura PL-Grid
- Gauss Centre for Supercomputing
- Academic Computer Centre Cyfronet, AGH University of Science and Technology