The spatial organization of membrane-bound ligands is thought to regulate receptor-mediated signaling. However, direct regulation of receptor function by nanoscale distribution of ligands has not yet been demonstrated, to our knowledge. We developed rationally designed DNA origami nanostructures modified with ligands at well-defined positions. Using these 'nanocalipers' to present ephrin ligands, we showed that the nanoscale spacing of ephrin-A5 directs the levels of EphA2 receptor activation in human breast cancer cells. Furthermore, we found that the nanoscale distribution of ephrin-A5 regulates the invasive properties of breast cancer cells. Our ligand nanocaliper approach has the potential to provide insight into the roles of ligand nanoscale spatial distribution in membrane receptor–mediated signaling.
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We thank U. Lendahl and his lab (Karolinska Institutet) for reagents and discussions, J. Avila-Cariño for help with the FACS experiments, O. Shupliakov for help with TEM, G. Bernardinelli for help with rendering ephrin-A5 and Y.-X. Zhao for help with initial experiments. This work was funded through grants from the Swedish Research Council to B.H. (repatriation grant 2010-6296 and project grant 2010-5060) and by the Strategic Research Program in Stem Cell Research and Regenerative Medicine at Karolinska Institutet (StratRegen), Sweden (A.I.T.). B.H. received startup funding from Carl Bennet AB, Karolinska Institutet and the Swedish Governmental Agency for Innovation Systems (Vinnova). V.L. and A.S. were supported by KID doctoral fellowships from the Karolinska Institutet, Sweden.
The authors declare no competing financial interests.
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Shaw, A., Lundin, V., Petrova, E. et al. Spatial control of membrane receptor function using ligand nanocalipers. Nat Methods 11, 841–846 (2014). https://doi.org/10.1038/nmeth.3025
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