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. Author manuscript; available in PMC: 2020 Jul 18.
Published in final edited form as: Cell Chem Biol. 2019 May 2;26(7):980–990.e8. doi: 10.1016/j.chembiol.2019.03.012

Figure 1 |. Workflow for identifying site-specific kinase-substrate interactions.

Figure 1 |

A, PhAXA and structure of ATP crosslinker probe 1. B, HEK293T cells transfected with WT or 3XFLAG phosphosite-to-Cys mutant 4E-BP1 constructs were lysed, treated with 1 or ATP and analyzed via Western blot. A minimal mass shift is observed due to the negligible size of FLAG-4E-BP1 relative to mTOR. C, PP242 (10,000–1 nM) and D, rapamycin (100 nM) inhibit the crosslinking of mTOR to the T46C 4E-BP1 in the presence of 1.