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The Ras Dimer Structure

preprint
submitted on 16.02.2021, 16:34 and posted on 17.02.2021, 13:12 by Till Rudack, Christian Teuber, Marvin Scherlo, Jörn Güldenhaupt, Jonas Schartner, Mathias Lübben, Johann P. Klare, Klaus Gerwert, Carsten Kötting
Oncogenic mutated Ras is a key player in cancer, but despite intense and expensive approaches its catalytic center seems undruggable. The Ras dimer interface is a possible alternative drug target. Dimerization at the membrane affects cell growth signal transduction. In vivo studies indicate that preventing dimerization of oncogenic mutated Ras inhibits uncontrolled cell growth. Conventional computational drug-screening approaches require a precise atomic dimer model as input to successfully access drug candidates. However, the proposed dimer structural models are controversial. Here, we provide a clear-cut experimentally validated N-Ras dimer structural model. We incorporated unnatural amino acids into Ras to enable the binding of labels at multiple positions via click chemistry. This labeling allowed for the determination of multiple distances of the membrane-bound Ras-dimer measured by fluorescence and electron paramagnetic resonance spectroscopy. In combination with protein-protein docking and biomolecular simulations we identified key residues for dimerization. Site‑directed mutations of these residues prevent dimer formation in our experiments, proving our dimer model to be correct. The presented dimer structure enables now computational drug-screening studies exploiting the Ras dimer interface as alternative drug target.

Funding

Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) Individual Research Grant GE 599/20-1 to KG

Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) Individual Research Grant KO 3813/1-1 to CK

Ministry for Culture and Science (MKW) of North Rhine-Westphalia (Germany) through grant 111.08.03.05-133974 to KG

Protein Research Unit Ruhr within Europe (PURE) funded by the Ministry of Innovation, Science and Research (MIWF) of North-Rhine Westphalia (Germany) to KG

History

Email Address of Submitting Author

till.rudack@rub.de

Institution

Ruhr University Bochum

Country

Germany

ORCID For Submitting Author

0000-0003-2693-9561

Declaration of Conflict of Interest

None

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