PXD078205-1

PXD078205 is an original dataset announced via ProteomeXchange.

Title	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.
HostingRepository	iProX
AnnounceDate	2026-05-09
AnnouncementXML	Submission_2026-05-13_02:25:25.509.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Xinnan Zheng
SpeciesList	scientific name: Mus musculus; NCBI TaxID: 10090;
ModificationList	No PTMs are included in the dataset
Instrument	timsTOF HT

Revision	Datetime	Status	ChangeLog Entry
0	2026-05-09 04:59:32	ID requested
⏵ 1	2026-05-13 02:25:26	announced

Zheng XN, Zeng JM, Huang HY, Zhao C, Ma SS, Feng L, Zhu H, Tian L, A click-compatible BmTyr platform for biotin-free proximity labeling and subcellular proteome profiling in primary T cells. Biochem Biophys Res Commun, 822():153918(2026) [pubmed]

submitter keyword: BmTyr, bioorthogonal proximity labeling, Click Chemistry, HiBiT Tag

Tian Lin
contact affiliation	Sun Yat-sen University Cancer Center
contact email	tianlin@sysucc.org.cn
lab head
Xinnan Zheng
contact affiliation	Sun Yat-sen University Cancer Center
contact email	zhengxn@sysucc.org.cn
dataset submitter

iProX dataset URI