Computationally designed sensors detect endogenous Ras activity and signaling effectors at subcellular resolution.

Autor: Zhang JZ; Department of Biochemistry, University of Washington, Seattle, WA, USA. jzz0428@uw.edu.; Institute for Protein Design, University of Washington, Seattle, WA, USA. jzz0428@uw.edu.; Howard Hughes Medical Institute, University of Washington, Seattle, WA, USA. jzz0428@uw.edu., Nguyen WH; Department of Biochemistry, University of Washington, Seattle, WA, USA., Greenwood N; Department of Biochemistry, University of Washington, Seattle, WA, USA.; Institute for Protein Design, University of Washington, Seattle, WA, USA., Rose JC; Department of Dermatology, Stanford University School of Medicine, Stanford, CA, USA., Ong SE; Department of Pharmacology, University of Washington, Seattle, WA, USA., Maly DJ; Department of Biochemistry, University of Washington, Seattle, WA, USA. djmaly@uw.edu.; Department of Chemistry, University of Washington, Seattle, WA, USA. djmaly@uw.edu., Baker D; Department of Biochemistry, University of Washington, Seattle, WA, USA. dbaker@uw.edu.; Institute for Protein Design, University of Washington, Seattle, WA, USA. dbaker@uw.edu.; Howard Hughes Medical Institute, University of Washington, Seattle, WA, USA. dbaker@uw.edu.
Jazyk: angličtina
Zdroj: Nature biotechnology [Nat Biotechnol] 2024 Dec; Vol. 42 (12), pp. 1888-1898. Date of Electronic Publication: 2024 Jan 25.
DOI: 10.1038/s41587-023-02107-w
Abstrakt: The utility of genetically encoded biosensors for sensing the activity of signaling proteins has been hampered by a lack of strategies for matching sensor sensitivity to the physiological concentration range of the target. Here we used computational protein design to generate intracellular sensors of Ras activity (LOCKR-based Sensor for Ras activity (Ras-LOCKR-S)) and proximity labelers of the Ras signaling environment (LOCKR-based, Ras activity-dependent Proximity Labeler (Ras-LOCKR-PL)). These tools allow the detection of endogenous Ras activity and labeling of the surrounding environment at subcellular resolution. Using these sensors in human cancer cell lines, we identified Ras-interacting proteins in oncogenic EML4-Alk granules and found that Src-Associated in Mitosis 68-kDa (SAM68) protein specifically enhances Ras activity in the granules. The ability to subcellularly localize endogenous Ras activity should deepen our understanding of Ras function in health and disease and may suggest potential therapeutic strategies.
Competing Interests: Competing interests: J.Z.Z., D.J.M. and D.B. are co-inventors on a provisional patent application (application number 63/380,884, submitted by the University of Washington) covering the biosensors described in this paper. The other authors declare no competing interests.
(© 2024. The Author(s).)
Databáze: MEDLINE
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