CPC C12N 9/16 (2013.01) [A61K 39/39558 (2013.01); C07K 14/435 (2013.01); C12Y 301/03048 (2013.01); A61K 38/00 (2013.01); C07K 2319/03 (2013.01); C07K 2319/61 (2013.01)] | 23 Claims |
1. A Src homology region 2 domain-containing phosphatase-2 (Shp2)-actuating protein (iSNAP) (Shp2-iSNAP) chimeric protein comprising in either: amino terminal to carboxy terminal order, or carboxy terminal to amino terminal order, modules comprising:
(a) a bi-phosphorylatable peptide comprising a bisphosphoryl tyrosine-based activation (BTAM) motif,
(b) a Fluorescent Protein (FP) Förster Resonance Energy Transfer (FRET) (FP FRET) pair comprising an enhanced cyan fluorescent protein (ECFP) motif and a YPet fluorochrome,
(c) a truncated Shp2 domain consisting of an N-Src Homology 2 (N-SH2) domain and a C-Src Homology 2 (C-SH2) domain,
wherein an unphosphorylated Shp2 domain binds or quenches the PTP domain or the kinase domain; and,
(d) a phosphatase (PTP) domain or a kinase domain,
and when the bi-phosphorylatable peptide, when phosphorylated, binds the N-SH2 domain and the C-SH2 domain and liberates or unquenches or activates the PTP domain or the kinase domain, and unquenching or activating of the PTP domain or the kinase domain causes or results in the emission of a detectable signal by the YPet fluorochrome.
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