LOVTRAP (a method that uses the Zdk polypeptide)
Zdk is a small peptide, stable in living cells, that binds selectively to the dark state of LOV2. LOV2 is anchored to the mitochondrial outer membrane. The protein of interest (POI) is attached to Zdk. The POI is sequestered at mitochondria in the dark and released when cells are irradiated.
The light-induced conformational change of the LOV domain is rapid (< 0.5 milliseconds). The half time for return to the dark state can be adjusted using point mutations (in our hands from 1.7 to 496 seconds, Curr. Protoc. Cell Biol., 73: 21.10.1-14, 2016). Rapid return times are useful for precise kinetic control. Slower return times enable continuous activation with only a brief pulse of light every few minutes.
References
Takano, T., Wu, M., Nakamuta, S., Naoki, H., Ishizawa, N., Namba, T., Watanabe, T., Xu, C., Hamaguchi, T., Yura, Y., Amano, M., Hahn, K.M. and Kaibuchi, K. Discovery of long-range inhibitory signaling to ensure 0.3 single axon formation. Nature Communications, 8(1): 33, 2017. PMC5484694. Online article | Free PMC article
Wang, W., Vilela, M., Winkler, A., Tarnawski, T., Schlichting, I., Yumerefendi, H., Kuhlman, B., Liu, R., Danuser, G., and Hahn, K.M. LOVTRAP, An Optogenetic System for Photo-induced Protein Dissociation. Nature Methods, 13(9): 755-8, 2016. PMC5137947. Online article | Free PMC article
Methods articles
Wang, H. and Hahn, K.M. LOVTRAP: a versatile method to control protein function with light. Curr. Protoc. Cell Biol., 73: 21.10.1 - 21.10.14, 2016. PMC5137945. Online article | Free PMC article
~ Updated 04/12/2021