PXD041013
PXD041013 is an original dataset announced via ProteomeXchange.
Dataset Summary
| Title | Comprehensive Characterization of Endogenous Phospholamban Proteoforms Enabled by Photocleavable Surfactant and Top-down Proteomics |
| Description | Top-down mass spectrometry (MS)-based proteomics has become a powerful tool for analyzing intact proteins and their associated post-translational modification (PTMs). Membrane proteins play critical roles in numerous cellular functions and represent the largest class of drug targets. However, top-down MS characterization of endogenous membrane proteins remains challenging, mainly due to their hydrophobicity and low expression level. Phospholamban (PLN) is a regulatory membrane protein in the sarcoplasmic reticulum and is essential in controlling cardiac contraction through calcium handling. Endogenously, the dynamic regulation of PLN's combinatorial PTM state has a significant influence on cardiac contractility and disease. Herein, we have developed a rapid and robust top-down proteomics method enabled by a photocleavable anionic surfactant, Azo, for the extraction and comprehensive characterization of endogenous PLN from cardiac tissue. We have employed a two-pronged top-down MS approach utilizing an online reversed-phase liquid chromatography tandem MS (LC-MS/MS) on a quadrupole time-of-flight MS and an ultrahigh-resolution Fourier-transform ion cyclotron resonance (FTICR) MS via direct infusion. We have comprehensively characterized the sequence and combinatorial PTMs of endogenous PLN, including S-palmitoylation and phosphorylations. S-palmitoylation was localized to Cys36 and for the first time with MS/MS, we have successfully localized the site-specific phosphorylation to Ser16 and Thr17 in human PLN extracted directly from heart tissue. We envision this top-down proteomics method will be crucial in its application to understanding PLN's role in cardiac contractility. |
| HostingRepository | MassIVE |
| AnnounceDate | 2023-08-11 |
| AnnouncementXML | Submission_2023-08-11_21:29:25.999.xml |
| DigitalObjectIdentifier | |
| ReviewLevel | Non peer-reviewed dataset |
| DatasetOrigin | Original dataset |
| RepositorySupport | Unsupported dataset by repository |
| PrimarySubmitter | Holden Rogers |
| SpeciesList | scientific name: Homo sapiens; common name: human; NCBI TaxID: 9606; |
| ModificationList | Phospho; S-palmitoyl-L-cysteine |
| Instrument | maXis; impact II; solariX; nanoACQUITY UPLC |
Dataset History
| Revision | Datetime | Status | ChangeLog Entry |
|---|---|---|---|
| 0 | 2023-03-21 14:17:09 | ID requested | |
| ⏵ 1 | 2023-08-11 21:29:26 | announced |
Publication List
| no publication |
Keyword List
| submitter keyword: Phospholamban, Top-down Proteomics, Azo |
Contact List
| Ying Ge | |
|---|---|
| contact affiliation | University of Wisconsin-Madison |
| contact email | ge2@wisc.edu |
| lab head | |
| Holden Rogers | |
| contact affiliation | University of Wisconsin-Madison |
| contact email | htrogers@wisc.edu |
| lab head | |
| Holden Rogers | |
| contact affiliation | University of Wisconsin-Madison |
| contact email | htrogers@wisc.edu |
| dataset submitter | |
Full Dataset Link List
| MassIVE dataset URI |
| Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://massive.ucsd.edu/MSV000091522/ |
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