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. Author manuscript; available in PMC: 2014 Apr 19.
Published in final edited form as: ACS Chem Biol. 2013 Jan 23;8(4):691–699. doi: 10.1021/cb300623a

Figure 4.

Figure 4

Catch-and-release of proteins labeled by the kinase-directed photo-crosslinker 4. a) Chemical structure of Probe 4. Kinase inhibitor 3 was converted into a photo-crosslinker by attaching a photo-activatable benzophenone moiety and a hexylchloride tag. b) Pulldown schematic for the catch-and-release of proteins photo-labeled by probe 4. Cell lysate was incubated with probe 4, followed by irradiation with UV light. The lysate was then successively incubated with ASH* and CLP-resin. After extensive washing, captured proteins were released from the resin by incubating with Ulp1*. A parallel experiment was performed in the presence of an active site competitor (dasatinib). c) Western blot analysis (anti-SRC) of the catch-and-release of the tyrosine kinase SRC. Photo-labeled SRC is captured by ASH* and a SRC/HaloTag complex is released from the beads with Ulp1*.