<<< Full experiment listing

PXD041013

PXD041013 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleComprehensive Characterization of Endogenous Phospholamban Proteoforms Enabled by Photocleavable Surfactant and Top-down Proteomics
DescriptionTop-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.
HostingRepositoryMassIVE
AnnounceDate2023-08-11
AnnouncementXMLSubmission_2023-08-11_21:29:25.999.xml
DigitalObjectIdentifier
ReviewLevelNon peer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterHolden Rogers
SpeciesList scientific name: Homo sapiens; common name: human; NCBI TaxID: 9606;
ModificationListPhospho; S-palmitoyl-L-cysteine
InstrumentmaXis; impact II; solariX; nanoACQUITY UPLC
Dataset History
RevisionDatetimeStatusChangeLog Entry
02023-03-21 14:17:09ID requested
12023-08-11 21:29:26announced
Publication List
no publication
Keyword List
submitter keyword: Phospholamban, Top-down Proteomics, Azo
Contact List
Ying Ge
contact affiliationUniversity of Wisconsin-Madison
contact emailge2@wisc.edu
lab head
Holden Rogers
contact affiliationUniversity of Wisconsin-Madison
contact emailhtrogers@wisc.edu
lab head
Holden Rogers
contact affiliationUniversity of Wisconsin-Madison
contact emailhtrogers@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/