SUMO is a post‐translational modifier critical for cell cycle progression and genome stability that plays a role in tumorigenesis, thus rendering SUMO‐specific enzymes potential pharmacological targets. However, the systematic generation of tools for the activity profiling of SUMO‐specific enzymes has proven challenging. We developed a diversifiable synthetic platform for SUMO‐based probes by using a direct linear synthesis method, which permits N‐ and C‐terminal labelling to incorporate dyes and reactive warheads, respectively. In this manner, activity‐based probes (ABPs) for SUMO‐1, SUMO‐2, and SUMO‐3‐specific proteases were generated and validated in cells using gel‐based assays and confocal microscopy. We further expanded our toolbox with the synthesis of a K11‐linked diSUMO‐2 probe to study the proteolytic cleavage of SUMO chains. Together, these ABPs demonstrate the versatility and specificity of our synthetic SUMO platform for in vitro and in vivo characterization of the SUMO protease family.

+More

direct linear synthesis method posttranslational modifier vitro and in vivo characterization tumorigenesis cell cycle progression sumobased probes k11linked disumo2 probe sumo1 sumo2 cterminal labelling gelbased assays n sumo3specific proteases activity profiling of sumospecific enzymes proteolytic cleavage of sumo chains diversifiable synthetic platform warheads sumo protease genome stability

Dr. Monique P. C. Mulder , Dr. Remco Merkx , Katharina F. Witting , Dharjath S. Hameed , Dris El Atmioui , Lindsey Lelieveld , Frauke Liebelt , Prof. Dr. Jacques Neefjes , Dr. Ilana Berlin , Dr. Alfred C. O. Vertegaal , Prof. Dr. Huib Ovaa,.Total Chemical Synthesis of SUMO and SUMO‐Based Probes for Profiling the Activity of SUMO‐Specific Proteases. 57 (57),8958-8962.

Dr. Monique P. C. Mulder , Dr. Remco Merkx , Katharina F. Witting , Dharjath S. Hameed , Dris El Atmioui , Lindsey Lelieveld , Frauke Liebelt , Prof. Dr. Jacques Neefjes , Dr. Ilana Berlin , Dr. Alfred C. O. Vertegaal , Prof. Dr. Huib Ovaa,"Total Chemical Synthesis of SUMO and SUMO‐Based Probes for Profiling the Activity of SUMO‐Specific Proteases" 57

Dr. Monique P. C. Mulder , Dr. Remco Merkx , Katharina F. Witting , Dharjath S. Hameed , Dris El Atmioui , Lindsey Lelieveld , Frauke Liebelt , Prof. Dr. Jacques Neefjes , Dr. Ilana Berlin , Dr. Alfred C. O. Vertegaal , Prof. Dr. Huib Ovaa,"Total Chemical Synthesis of SUMO and SUMO‐Based Probes for Profiling the Activity of SUMO‐Specific Proteases"