PXD052354

PXD052354 is an original dataset announced via ProteomeXchange.

Dataset Summary

Title	Online Mixed-Bed Ion Exchange Chromatography for Native Top-Down Proteomics of Complex Mixtures
Description	Native top-down mass spectrometry (nTDMS) allows characterization of protein structure and noncovalent interactions with simultaneous sequence mapping and proteoform characterization. The majority of nTDMS studies utilize purified recombinant proteins, with significant challenges hindering application to endogenous systems. To perform native top-down proteomics (nTDP), where endogenous proteins from complex biological systems are analyzed by nTDMS, it is essential to separate proteins under nondenaturing conditions. However, it remains difficult to achieve high resolution and MS-compatibility with online chromatography while preserving protein tertiary structure and noncovalent interactions. Herein, we report the use of online mixed-bed ion exchange chromatography (IEC) to enable separation of endogenous proteins from complex mixtures under nondenaturing conditions, preserving noncovalent interactions for nTDP analysis. We have successfully detected large proteins (>146 kDa) and identified endogenous metal-binding and oligomeric protein complexes in human heart tissue lysate. The use of a mixed-bed stationary phase allowed retention and elution of proteins over a wide range of isoelectric points without altering the sample or mobile phase pH. Overall, our method provides a simple online IEC-MS platform that can effectively separate proteins from complex mixtures under nondenaturing conditions and preserve higher-order structure for nTDP applications.
HostingRepository	MassIVE
AnnounceDate	2025-04-10
AnnouncementXML	Submission_2025-04-10_12:49:06.250.xml
DigitalObjectIdentifier
ReviewLevel	Non peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Matthew
SpeciesList	scientific name: Homo sapiens; common name: human; NCBI TaxID: 9606;
ModificationList	Phospho
Instrument	maXis II

Dataset History

Revision	Datetime	Status	ChangeLog Entry
0	2024-05-17 09:54:19	ID requested
⏵ 1	2025-04-10 12:49:06	announced

Publication List

no publication

no publication

Keyword List

submitter keyword: native, top-down proteomics, ion-exchange chromatography

submitter keyword: native, top-down proteomics, ion-exchange chromatography

Contact List

Ying Ge
contact affiliation	University of Wisconsin-Madison
contact email	ying.ge@wisc.edu
lab head
Matthew
contact affiliation	UW-Madison
contact email	msfischer5@wisc.edu
dataset submitter

Full Dataset Link List

MassIVE dataset URI
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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-ftp.ucsd.edu/v07/MSV000094787/