Scheme 2.
Synthesis of the seven isopeptide‐linked diUb FRET pairs. a) 10, PyBOP, DIPEA, NMP; b) 20 % HFIP/CH2Cl2; c) HCl⋅H‐Gly‐S(CH2)2CO2Me, EDC, HOBt, CH2Cl2; d) TFA/H2O/phenol/iPr3SiH (90.5:5:2.5:2); e) TAMRA, PyBOP, DIPEA, NMP; f) TFA/H2O/phenol/iPr3SiH (90.5:5:2.5:2); g) MPAA, 6 m Gnd⋅HCl, pH 7.2; h) TCEP, GSH, VA‐044, 6 m Gnd⋅HCl, pH 7.0.