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Rationally improving LOV domain–based photoswitches

Abstract

Genetically encoded protein photosensors are promising tools for engineering optical control of cellular behavior; we are only beginning to understand how to couple these light detectors to effectors of choice. Here we report a method that increases the dynamic range of an artificial photoswitch based on the LOV2 domain of Avena sativa phototropin 1 (AsLOV2). This approach can potentially be used to improve many AsLOV2-based photoswitches.

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Figure 1: Conformational and binding equilibria in AsLOV2 and LovTAP.
Figure 2: Mutational stabilization of the LOV-Jα association and its effect on DNA-binding activity in LovTAP.

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Acknowledgements

We thank E. Munro and A. Möglich for critical reading of the manuscript; S. Crosson and T. Pan for sharing reagents and equipment and members of the Sosnick, Gardner and Rosen laboratories for helpful discussions. This work was supported by grants from the US National Institutes of Health (R01 GM081875 to K.H.G., GM55694 to T.R.S. and GM088668 to T.R.S. and M. Glotzer), the Robert A. Welch Foundation (I-1424 to K.H.G. and I-1544 to M.K.R.) and the Chicago Biomedical Consortium with support from The Searle Funds at The Chicago Community Trust (to T.R.S., M. Glotzer and E. Weiss).

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D.S. designed and performed experiments, analyzed data and wrote the paper; X.Y. designed and performed experiments, analyzed data and wrote the paper; G.G. performed experiments and analyzed data; M.K.R. designed experiments, analyzed data and wrote the paper; K.H.G. designed experiments, analyzed data and wrote the paper; T.R.S. designed experiments, analyzed data and wrote the paper.

Corresponding authors

Correspondence to Michael K Rosen or Kevin H Gardner or Tobin R Sosnick.

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The authors declare no competing financial interests.

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Strickland, D., Yao, X., Gawlak, G. et al. Rationally improving LOV domain–based photoswitches. Nat Methods 7, 623–626 (2010). https://doi.org/10.1038/nmeth.1473

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