E3 activity-based proteomic profiling of HEK293 cells treated with ATPgammaS

RNF213 is novel transthiolating E3 ligase that is activated by ATP. To determine whether cellular RNF213 is activated by elevated ATP levels, we electroplated an E3 activity-based probe into HEK293 cells stably expressing RNF213 in a knockout background. DIA analysis was performed in cells that were coelectroporated with the poorly hydrolysable ATP analog, ATPgS, or buffer control.