PXD045682
PXD045682 is an original dataset announced via ProteomeXchange.
Dataset Summary
Title | [HPP-2.0] A tyrosine phosphoproteome analysis approach enabled by selective dephosphorylation with protein tyrosine phosphatase |
Description | Protein tyrosine phosphorylation (pTyr) plays a prominent role in signal transduction and regulation in all eukaryotic cells. In conventional immunoaffinity purification (IP) methods, phosphotyrosine peptides are isolated from the digest of cellular protein extracts with a phosphotyrosine-specific antibody and are identified by tandem mass spectrometry. However, the low sensitivity, poor reproducibility and cost-prohibitive are universal concerns for IP approaches. In this study, we presented an antibody-free approach to identify phosphortyrosine peptides by using protein tyrosine phosphatase (PTP). It was found that most of PTPs including PTP1B, TCPTP and SHP1 can efficiently and selectively dephosphorylated phosphotyrosine peptides. We then designed a workflow by combining two Ti4+-IMAC-based phosphopeptide enrichment steps with PTP catalyzed dephosphorylation for tyrosine phosphoproteomics analysis. This workflow was firstly validated by selectively detection of phosphotyrosine peptides from semi-complex sample and then applied to analyze the tyrosine phosphoproteome of Jurkat T cells. Around 1000 putative former phosphotyrosine peptides were identified from less than 500 μg of cell lysate. The tyrosine phosphosites on majority of these peptides could be unambiguously determined for over 70% of them possess only one tyrosine residue. It was also found that the tyrosine sites identified by this method was highly complementary to these identified by SH2 superbinder based method. Therefore, the combination of Ti4+-IMAC enrichment with PTP dephosphorylation provides an alternative and cost-effective approach for tyrosine phosphoproteomics analysis. |
HostingRepository | iProX |
AnnounceDate | 2023-09-26 |
AnnouncementXML | Submission_2024-05-21_00:51:22.773.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Zheng Fang |
SpeciesList | scientific name: Homo sapiens; NCBI TaxID: 9606; |
ModificationList | No PTMs are included in the dataset |
Instrument | Q Exactive HF |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
---|---|---|---|
0 | 2023-09-25 20:22:08 | ID requested | |
1 | 2023-09-25 20:22:16 | announced | |
⏵ 2 | 2024-05-21 00:51:23 | announced | 2024-05-21: Update information. |
Publication List
Liu X, Dong M, Yao Y, Wang Y, Mao J, Hu L, Yao L, Ye M, A Tyrosine Phosphoproteome Analysis Approach Enabled by Selective Dephosphorylation with Protein Tyrosine Phosphatase. Anal Chem, 94(10):4155-4164(2022) [pubmed] |
Keyword List
submitter keyword: Phospho-Proteome |
Contact List
Mingliang Ye | |
---|---|
contact affiliation | Dalian Institute of Chemical Physics, Chinese Academy of Sciences |
contact email | mingliang@dicp.ac.cn |
lab head | |
Zheng Fang | |
contact affiliation | Dalian Institute of Chemical Physics, Chinese Academy of Sciences |
contact email | zhengfang@dicp.ac.cn |
dataset submitter |
Full Dataset Link List
iProX dataset URI |