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PXD041013-1

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
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