PXD046150

PXD046150 is an original dataset announced via ProteomeXchange.

Title	Cellular protein painting for structural and binding sites analysis via lysine reactivity profiling with o-phthalaldehyde
Description	The three-dimensional structure and the molecular interaction of proteins determine their roles in many cellular processes. Chemical protein painting with protein mass spectrometry can identify changes in structural conformations and molecular interactions of proteins including their binding sites. Nevertheless, most current protein painting techniques identified protein targets and binding sites of drugs in vitro using cell lysate or purified protein. Here, we screened 11 kinds of membrane-permeable lysine-reactive chemical probes for intracellular covalent labeling of endogenous proteins, which reveals ortho-phthalaldehyde (OPA) as the most reactive probe in intracellular environment. An MS workflow was developed and coupled with a new data analysis strategy termed RAPID (Reactive Amino acid Profiling by Inverse Detection) to enhance detection sensitivity. RAPID-OPA successfully identified structural change induced by allosteric drug TEPP-46 on its target protein PKM2, and was applied to profile conformation change of the proteome occurring in cells during thermal denaturation. Application of RAPID-OPA on cells treated with three model drugs geldanamycin, selumetinib, and staurosporine successfully revealed its binding sites on target proteins. Thus, RAPID-OPA for cellular protein painting permits the identification of ligand-binding sites and detection of protein structural changes occurring in cells.
HostingRepository	iProX
AnnounceDate	2023-10-16
AnnouncementXML	Submission_2024-05-20_20:18:03.666.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Zhenxiang Zheng
SpeciesList	scientific name: Homo sapiens; NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive; Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2023-10-15 22:32:31	ID requested
1	2023-10-31 20:05:29	announced
⏵ 2	2024-05-20 20:18:04	announced	2024-05-21: Update information.

Zheng Z, Zeng Y, Lai K, Liao B, Li P, Tan CSH, -phthalaldehyde. Chem Sci, 15(16):6064-6075(2024) [pubmed]

submitter keyword: Protein structural conformation, binding site, chemical probe, Proteomics

Chris Soon Heng Tan
contact affiliation	Southern University of Science and Technology
contact email	christan@sustech.edu.cn
lab head
Zhenxiang Zheng
contact affiliation	Southern University of Science and Technology
contact email	12031007@mail.sustech.edu.cn
dataset submitter

iProX dataset URI