Glycine Fluoromethylketones as SENP‐Specific Activity Based Probes

Abstract

We report here the synthesis and biochemical properties of a new peptidyl activity-based probe 1 for SUMO proteases, SENPs. The activity-based probe has at its C terminus a glycine-derived fluoromethylketone moiety as a reactive group designed to target the active-site cysteine of SENPs. Based on a study of the interactions between SENPs and SUMOs, we introduced further design elements that allow the activity-based probe to selectively target SENPs at low micromolar to high nanomolar concentrations. Moreover, 1 out-competes SUMO1 from the reversible SUMO1-SENP1 complex, thus suggesting that 1 and SUMO1 share a common binding site on SENP1.

Sulfonyl Fluoride Analogues as Activity‐Based Probes for Serine Proteases

“…The metalloproteases have been adapted to ABPP technologies by incorporating photo‐crosslinking benzophenones within known noncovalent inhibitor scaffolds 5. A wide variety of chemical probes incorporating acyloxymethyl ketones,6 epoxysuccinates,7 halomethylketones,8 and vinyl sulfones9 have been applied to profile numerous members of the cysteine protease family. However, compared to the armory of ABPs available for cysteine proteases, ABPs for the serine protease family are poorly represented, mostly due to the lower reactivity of the serine nucleophile relative to its cysteine counterpart.…”

Total Chemical Synthesis of SUMO and SUMO‐Based Probes for Profiling the Activity of SUMO‐Specific Proteases

“…It has been established that many tumours rely on a functional SUMO pathway, which makes SUMO‐related processes potential anti‐cancer drug targets . However, the molecular mechanisms underlying the role of SUMO modification in these processes remain poorly understood, in part due to the slow development of activity‐based reagents . In contrast, research in the ubiquitin field has accelerated due to the convenient chemical synthesis of ubiquitin, which has given access to assay reagents and probes Therefore, a reliable route towards SUMO‐based conjugates is urgently needed to provide the reagents required to visualize and understand the biology of reversible SUMOylation.…”

Total Chemical Synthesis of SUMO and SUMO‐Based Probes for Profiling the Activity of SUMO‐Specific Proteases

“…It has been established that many tumours rely on a functional SUMO pathway, which makes SUMO‐related processes potential anti‐cancer drug targets 8. However, the molecular mechanisms underlying the role of SUMO modification in these processes remain poorly understood, in part due to the slow development of activity‐based reagents 9. In contrast, research in the ubiquitin field has accelerated due to the convenient chemical synthesis of ubiquitin,10 which has given access to assay reagents and probes11, 12, 13, 14 Therefore, a reliable route towards SUMO‐based conjugates is urgently needed to provide the reagents required to visualize and understand the biology of reversible SUMOylation.…”