{"database":"iProX","file_versions":[],"scores":null,"additional":{"omics_type":["Proteomics"],"submitter":["Xu Li"],"species":["Homo Sapiens"],"full_dataset_link":["http://www.iprox.org/page/project.html?id=IPX0018073000"],"submitter_email":["lixuqd@qdu.edu.cn"],"submitter_affiliation":["Qingdao University"],"sample_protocol":[""],"repository":["iProX"],"data_protocol":[""],"additional_accession":[]},"is_claimable":false,"name":"Activity-Based Ubiquitin-α-Amidomethyl Acrylate Probe for Profiling of Ubiquitinating Enzymes and Deubiquitinases","description":"Activity-based ubiquitin probes have emerged as powerful tools for dissecting protein ubiquitination. In this work, we discovered that an α-amidomethyl substituted acrylate electrophile exhibits markedly enhanced reactivity. Based on this finding, we designed and synthesized a new activity-based probe, Ubiquitin-α-Amidomethyl Acrylate (Ub-AMA). In vitro biochemical assays demonstrated that Ub-AMA exhibits broad crosslinking capability across the entire ubiquitin regulatory enzyme system—including E1, E2, and E3 enzymes as well as DUBs. Compared to the classic Ub-VME probe, Ub-AMA achieves significantly higher labeling efficiency for E2 enzymes. Finally, proteomic profiling in cell lysates confirmed that Ub-AMA can capture E1, E2, and E3 enzymes as well as DUBs in a complex biological environment. In summary, the Ub-AMA probe developed here provides a new tool for in depth analysis of the dynamic regulatory mechanisms of ubiquitination. Moreover, this work offers valuable insights for the design of other activity based probes based on α,β-unsaturated carbonyl scaffolds.","dates":{"publication":"Tue Jun 30 00:00:00 GMT+01:00 2026"},"accession":"PXD080401","cross_references":{"TAXONOMY":["9606"]}}