Direct Modulators of K-Ras–Membrane Interactions
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1.
University of California, San Francisco
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2.
California Institute of Technology
Abstract
Protein–membrane interactions (PMIs) are ubiquitous in cellular signaling. Initial steps of signal transduction cascades often rely on transient and dynamic interactions with the inner plasma membrane leaflet to populate and regulate signaling hotspots. Methods to target and modulate these interactions could yield attractive tool compounds and drug candidates. Here, we demonstrate that the conjugation of a medium-chain lipid tail to the covalent K-Ras(G12C) binder MRTX849 at a solvent-exposed site enables such direct modulation of PMIs. The conjugated lipid tail interacts with the tethered membrane and changes the relative membrane orientation and conformation of K-Ras(G12C), as shown by molecular dynamics (MD) simulation-supported NMR studies. In cells, this PMI modulation restricts the lateral mobility of K-Ras(G12C) and disrupts nanoclusters. The described strategy could be broadly applicable to selectively modulate transient PMIs.
Copyright and License
Copyright © 2023 The Authors. Published by American Chemical Society
Acknowledgement
J.M. thanks the NCI for a K99/R00 award (K99CA277358). K.M.S. thanks NIH grant 5R01CA244550 and the Samuel Waxman Cancer Research Foundation. The authors thank J. O’Connell from B. Shoichet’s lab for assistance with the dynamic light scattering (DLS) measurement. The authors wish to acknowledge C. J. DeHart, J.-P. Denson, P. H. Frank, M. Hong, S. Messing, A. Mitchell, N. Ramakrishnan, W. Burgan, K. Powell, and T. Taylor for cloning, protein expression, protein purification, cell line production, and electrospray ionization mass spectroscopy. The authors thank J. B. Combs, P. Pfaff, and D. M. Peacock for the critical review of the manuscript. The authors also thank Q. Zheng for providing optimized conditions for the Cbz-deprotection step. This project has been funded in whole or in part with Federal funds from the National Cancer Institute, National Institutes of Health, under Contract No. 75N91019D00024. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services nor does the mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government. This study made use of the National Magnetic Resonance Facility at Madison, which is supported by NIH grants P41GM136463 and R24GM141526.
Conflict of Interest
The authors declare the following competing financial interest(s): K.M.S. and J.M. are inventors on patents owned by UCSF covering K-Ras targeting small molecules. K.M.S. has consulting agreements for the following companies, which involve monetary and/or stock compensation: Revolution Medicines, Black Diamond Therapeutics, BridGene Biosciences, Denali Therapeutics, Dice Molecules, eFFECTOR Therapeutics, Erasca, Genentech/Roche, Janssen Pharmaceuticals, Kumquat Biosciences, Kura Oncology, Mitokinin, Nested, Type6 Therapeutics, Venthera, Wellspring Biosciences (Araxes Pharma), Turning Point, Ikena, Initial Therapeutics, Vevo and BioTheryX.
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Additional details
- National Cancer Institute
- K99CA277358
- Samuel Waxman Cancer Research Foundation
- National Institutes of Health
- 5R01CA244550
- National Institutes of Health
- 75N91019D00024
- National Institutes of Health
- P41GM136463
- National Institutes of Health
- R24GM141526
- Publication Status
- Published