BRET-based RAS Biosensors That Show a Novel Small Molecule is an Inhibitor of RAS-effector Protein-protein Interactions

Overview

Journal Elife

Specialty Biology

Date 2018 Jul 11

PMID 29989546

Citations 38

Authors

Nicolas Bery

Abimael Cruz-Migoni

Carole Jr Bataille

Camilo E Quevedo

Hanna Tulmin

Ami Miller

Angela Russell

Simon Ev Phillips

Stephen B Carr

Terence H Rabbitts

Affiliations

Soon will be listed here.

Abstract

The RAS family of proteins is amongst the most highly mutated in human cancers and has so far eluded drug therapy. Currently, much effort is being made to discover mutant RAS inhibitors and in vitro screening for RAS-binding drugs must be followed by cell-based assays. Here, we have developed a robust set of bioluminescence resonance energy transfer (BRET)-based RAS biosensors that enable monitoring of RAS-effector interaction inhibition in living cells. These include KRAS, HRAS and NRAS and a variety of different mutations that mirror those found in human cancers with the major RAS effectors such as CRAF, PI3K and RALGDS. We highlighted the utility of these RAS biosensors by showing a RAS-binding compound is a potent pan-RAS-effector interactions inhibitor in cells. The RAS biosensors represent a useful tool to investigate and characterize the potency of anti-RAS inhibitors in cells and more generally any RAS protein-protein interaction (PPI) in cells.

Citing Articles

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References

Svensson O, Malbet-Monaco S, Popov A, Nurizzo D, Bowler M . Fully automatic characterization and data collection from crystals of biological macromolecules. Acta Crystallogr D Biol Crystallogr. 2015; 71(Pt 8):1757-67. PMC: 4528805. DOI: 10.1107/S1399004715011918. View

Hunter J, Manandhar A, Carrasco M, Gurbani D, Gondi S, Westover K . Biochemical and Structural Analysis of Common Cancer-Associated KRAS Mutations. Mol Cancer Res. 2015; 13(9):1325-35. DOI: 10.1158/1541-7786.MCR-15-0203. View

Prior I, Lewis P, Mattos C . A comprehensive survey of Ras mutations in cancer. Cancer Res. 2012; 72(10):2457-67. PMC: 3354961. DOI: 10.1158/0008-5472.CAN-11-2612. View

Cox A, Fesik S, Kimmelman A, Luo J, Der C . Drugging the undruggable RAS: Mission possible?. Nat Rev Drug Discov. 2014; 13(11):828-51. PMC: 4355017. DOI: 10.1038/nrd4389. View

Emsley P, Lohkamp B, Scott W, Cowtan K . Features and development of Coot. Acta Crystallogr D Biol Crystallogr. 2010; 66(Pt 4):486-501. PMC: 2852313. DOI: 10.1107/S0907444910007493. View

Tanaka T, Sewell H, Waters S, Phillips S, Rabbitts T . Single domain intracellular antibodies from diverse libraries: emphasizing dual functions of LMO2 protein interactions using a single VH domain. J Biol Chem. 2010; 286(5):3707-16. PMC: 3030373. DOI: 10.1074/jbc.M110.188193. View

McCoy A, Grosse-Kunstleve R, Adams P, Winn M, Storoni L, Read R . Phaser crystallographic software. J Appl Crystallogr. 2009; 40(Pt 4):658-674. PMC: 2483472. DOI: 10.1107/S0021889807021206. View

Chen V, Arendall 3rd W, Headd J, Keedy D, Immormino R, Kapral G . MolProbity: all-atom structure validation for macromolecular crystallography. Acta Crystallogr D Biol Crystallogr. 2010; 66(Pt 1):12-21. PMC: 2803126. DOI: 10.1107/S0907444909042073. View

Zhang B, Li S, Harbrecht B . Akt-mediated signaling is induced by cytokines and cyclic adenosine monophosphate and suppresses hepatocyte inducible nitric oxide synthase expression independent of MAPK P44/42. Biochim Biophys Acta. 2010; 1813(1):73-9. PMC: 3014387. DOI: 10.1016/j.bbamcr.2010.10.001. View

10.

Burgering B, Medema R, Van Weeren P, Tertoolen L, Bos J . Epidermal growth factor induces phosphorylation of extracellular signal-regulated kinase 2 via multiple pathways. Mol Cell Biol. 1993; 13(12):7248-56. PMC: 364795. DOI: 10.1128/mcb.13.12.7248-7256.1993. View

11.

Pfleger K, Seeber R, Eidne K . Bioluminescence resonance energy transfer (BRET) for the real-time detection of protein-protein interactions. Nat Protoc. 2007; 1(1):337-45. DOI: 10.1038/nprot.2006.52. View

12.

Bacart J, Corbel C, Jockers R, Bach S, Couturier C . The BRET technology and its application to screening assays. Biotechnol J. 2008; 3(3):311-24. DOI: 10.1002/biot.200700222. View

13.

Assi S, Tanaka T, Rabbitts T, Fernandez-Fuentes N . PCRPi: Presaging Critical Residues in Protein interfaces, a new computational tool to chart hot spots in protein interfaces. Nucleic Acids Res. 2009; 38(6):e86. PMC: 2847225. DOI: 10.1093/nar/gkp1158. View

14.

Waldmann H, Karaguni I, Carpintero M, Gourzoulidou E, Herrmann C, Brockmann C . Sulindac-derived Ras pathway inhibitors target the Ras-Raf interaction and downstream effectors in the Ras pathway. Angew Chem Int Ed Engl. 2004; 43(4):454-8. DOI: 10.1002/anie.200353089. View

15.

Lito P, Solomon M, Li L, Hansen R, Rosen N . Allele-specific inhibitors inactivate mutant KRAS G12C by a trapping mechanism. Science. 2016; 351(6273):604-8. PMC: 4955282. DOI: 10.1126/science.aad6204. View

16.

De A, Loening A, Gambhir S . An improved bioluminescence resonance energy transfer strategy for imaging intracellular events in single cells and living subjects. Cancer Res. 2007; 67(15):7175-83. PMC: 4161127. DOI: 10.1158/0008-5472.CAN-06-4623. View

17.

Spiegel J, Cromm P, Zimmermann G, Grossmann T, Waldmann H . Small-molecule modulation of Ras signaling. Nat Chem Biol. 2014; 10(8):613-22. DOI: 10.1038/nchembio.1560. View

18.

Wennerberg K, Rossman K, Der C . The Ras superfamily at a glance. J Cell Sci. 2005; 118(Pt 5):843-6. DOI: 10.1242/jcs.01660. View

19.

Shima F, Yoshikawa Y, Ye M, Araki M, Matsumoto S, Liao J . In silico discovery of small-molecule Ras inhibitors that display antitumor activity by blocking the Ras-effector interaction. Proc Natl Acad Sci U S A. 2013; 110(20):8182-7. PMC: 3657810. DOI: 10.1073/pnas.1217730110. View

20.

Kleeff J, Korc M, Apte M, La Vecchia C, Johnson C, Biankin A . Pancreatic cancer. Nat Rev Dis Primers. 2016; 2:16022. DOI: 10.1038/nrdp.2016.22. View