PXD019368-2

PXD019368 is an original dataset announced via ProteomeXchange.

Title	Top-down Proteomics of Endogenous Membrane Proteins Enabled by A Cloud Point Enrichment and Multidimensional Liquid Chromatography-Mass Spectrometry Top-down Proteomics of Endogenous Membrane Proteins Enabled by A Cloud Point Enrichment and Multidimensional Liquid Chromatography-Mass Spectrometry
Description	Top-down proteomics has emerged as a powerful strategy to characterize proteins in biological systems. Nevertheless, the analysis of endogenous membrane proteins is challenging due to their low solubility, low abundance, and the complexity of the membrane subproteome. Here, we report a simple, effective enrichment and separation strategy for top-down proteomics of endogenous membrane proteins enabled by cloud-point enrichment and multidimensional liquid chromatography coupled to high-resolution mass spectrometry (MS). The cloud point extraction efficiently enriched membrane proteins using a single extraction, eliminating the need for time-consuming ultracentrifugation steps. Subsequently, size-exclusion chromatography with an MS-compatible mobile phase was used to fractionate intact proteins (6-115 kDa). Fractions were separated further by reversed-phase liquid chromatography coupled with MS for protein characterization. We applied this method to human embryonic kidney cells and cardiac tissue, enabling the identification of 188 and 124 endogenous integral membrane proteins respectively, some with as many as 19 transmembrane domains.
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_05:15:51.578.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	kyle brown
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	phosphorylated residue; monohydroxylated residue; acetylated residue
Instrument	maXis

Revision	Datetime	Status	ChangeLog Entry
0	2020-05-25 04:00:51	ID requested
1	2020-11-30 00:15:13	announced
⏵ 2	2024-10-22 05:15:52	announced	2024-10-22: Updated project metadata.

10.1021/acs.analchem.0c02533;

Brown KA, Tucholski T, Alpert AJ, Eken C, Wesemann L, Kyrvasilis A, Jin S, Ge Y, Top-Down Proteomics of Endogenous Membrane Proteins Enabled by Cloud Point Enrichment and Multidimensional Liquid Chromatography-Mass Spectrometry. Anal Chem, 92(24):15726-15735(2020) [pubmed]

submitter keyword: Membrane protein, top-down proteomics, cloud point

Ying Ge
contact affiliation	University of Wisconsin-Madison
contact email	ying.ge@wisc.edu
lab head
kyle brown
contact affiliation	uw madison
contact email	kbrown33@wisc.edu
dataset submitter

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://ftp.pride.ebi.ac.uk/pride/data/archive/2020/11/PXD019368

PRIDE project URI

• PRIDE
  1. PXD019368
     1. Label: PRIDE project
     2. Name: Top-down Proteomics of Endogenous Membrane Proteins Enabled by A Cloud Point Enrichment and Multidimensional Liquid Chromatography-Mass Spectrometry Top-down Proteomics of Endogenous Membrane Proteins Enabled by A Cloud Point Enrichment and Multidimensional Liquid Chromatography-Mass Spectrometry