{"database":"iProX","file_versions":[],"scores":null,"additional":{"omics_type":["Proteomics"],"submitter":["Tian Lin"],"species":["Mus Musculus"],"full_dataset_link":["http://www.iprox.org/page/project.html?id=IPX0016748000"],"submitter_email":["tianlin@sysucc.org.cn"],"submitter_affiliation":["Sun Yat-sen University Cancer Center"],"sample_protocol":[""],"repository":["iProX"],"data_protocol":[""],"pubmed_abstract":["Proximity labeling has revolutionized the study of dynamic subcellular proteomes by enabling the capture of transient protein interactions within living cells, yet the application of existing platforms to hard-to-transfect primary cells remains challenging. Here, we leverage a bioorthogonal proximity labeling platform based on the copper-dependent tyrosinase BmTyr to profile subcellular proteomes in primary T cells. This system catalyzes the subcellular incorporation of an alkyne-phenol probe, enabling subsequent click-compatible conjugation to versatile azide-bearing tags for fluorescence imaging and affinity enrichment for mass spectrometry. To expand the proximity labeling toolkit, we developed a custom azide-HiBiT/His tag mixture, which enables direct, antibody-independent validation using the same alkyne-phenol labeling chemistry, coupled with efficient elution and ultrasensitive chemiluminescent detection for low-input samples. Applying this platform to primary T cells not only validated known nuclear components of the TNFα signaling pathway but also revealed a previously unappreciated chromatin-associated localization for NKAP, providing new mechanistic insight beyond its previously described nuclear translocation. Collectively, our work establishes a powerful and flexible tool for sensitive, context-specific proteomic mapping in challenging physiological systems."],"pubmed_title":["A click-compatible BmTyr platform for biotin-free proximity labeling and subcellular proteome profiling in primary T cells."],"pubmed_authors":["Zheng Xin-Nan XN, Zeng Jing-Min JM, Huang Han-Ying HY, Zhao Chuang C, Ma Sheng-Suo SS, Feng Lin L, Zhu Hao H, Tian Lin L"],"additional_accession":[]},"is_claimable":false,"name":"A click-compatible BmTyr platform for biotin-free proximity labeling and subcellular proteome profiling in primary T cells","description":"Proximity labeling has revolutionized the study of dynamic subcellular proteomes by enabling the capture of transient protein interactions within living cells, yet the application of existing platforms to hard-to-transfect primary cells remains challenging. Here, we leverage a bioorthogonal proximity labeling platform based on the copper-dependent tyrosinase BmTyr to profile subcellular proteomes in primary T cells. This system catalyzes the subcellular incorporation of an alkyne-phenol probe, enabling subsequent click-compatible conjugation to versatile azide-bearing tags for fluorescence imaging and affinity enrichment for mass spectrometry. To expand the proximity labeling toolkit, we developed a custom azide-HiBiT/His tag mixture, which enables direct, antibody-independent validation using the same alkyne-phenol labeling chemistry, coupled with efficient elution and ultrasensitive chemiluminescent detection for low-input samples. Applying this platform to primary T cells not only validated known nuclear components of the TNFα signaling pathway but also revealed a previously unappreciated chromatin-associated localization for NKAP, providing new mechanistic insight beyond its previously described nuclear translocation. Collectively, our work establishes a powerful and flexible tool for sensitive, context-specific proteomic mapping in challenging physiological systems.","dates":{"publication":"Sat May 09 00:00:00 BST 2026"},"accession":"PXD078205","cross_references":{"TAXONOMY":["10090"],"pubmed":["42119177"]}}