hahn lab

dye-AR / endogenous

(LEFT) This Cdc42 biosensor was constructed from a fragment of WASP that binds selectively to activated Cdc42 (due to the green CRIB domain). The WASP fragment was derivatized with an environment-sensing dye that changed fluorescence when the biosensor bound Cdc42. Dye at position 271 (red) was in a hydrophobic pocket formed by amino acids from both WASP (blue) and Cdc42 (orange). (RIGHT) A library of FN3 monobodies is screened to find a library member with the appropriate binding selectivity and affinity for the targeted protein state. The library is based on a uniform scaffold stable in living cells and suitable for conversion to biosensors. The appropriate library member is derivatized with an environmentally sensitive dye to report target binding. Here, binding to active Src family kinases leads to increased fluorescence from a merocyanine dye.


Konze, K.D., Ma, A., Li, F., Barsyte-Lovejoy, D., Parton, T., MacNevin, C.J., Liu, F., Gao, C., Huang, X.P., Kuznetsova, E., Rougie, M., Jiang, A., Pattenden, S.G., Norris, J.L., James, L.I., Roth, B.L., Brown, P.J., Frye, S.V., Arrowsmith, C.H., Hahn, K.M., Wang, G.G., Vedadi, M. and Jin, J. An orally bioavailable chemical probe of the lysine methyltransferases EZH2 and EZH1. ACS Chem. Biol., 8(6): 1324–1334, 2013. PMC3773059. Online article | Free PMC article

Gulyani, A., Vitriol, E., Allen, R., Wu, J., Gremyachinskiy, D., Lewis, S., Dewar, B., Graves, L.M., Kay, B.K., Kuhlman, B., Elston T. and Hahn, K.M.  A biosensor generated via high-throughput screening quantifies cell edge Src dynamics. Nature Chem. Biol., 7: 437-444, 2011. PMC3135387. Online article | Free PMC article

Nalbant, P., L. Hodgson, V. Kraynov, A. Toutchkine, K.M. Hahn. Activation of Endogenous Cdc42 Visualized in Living Cells. Science, 305:1615-1619, 2004. Online article | PDF

Methods articles

Hodgson, L., Pertz, O., and Hahn, K.M. Design and Optimization of genetically encoded fluorescent biosensors: GTPase biosensors. Methods Cell Biol, 85:63-81, 2008. [Link downloads PDF] Online article

Hodgson, L. Nalbant, P. Shen, F., and Hahn, K. Imaging and photobleach correction of Mero-CBD, sensor of endogenous Cdc42 activation. Methods Enzymol., 406:140-156, 2006. Online article | PDF


~ Updated 04/08/2

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