PXD041013-1

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

no publication

Keyword List

submitter keyword: Phospholamban, Top-down Proteomics, Azo

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

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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/