PXD040309-2

PXD040309 is an original dataset announced via ProteomeXchange.

Title	Proteomic profiling of centrosomes across multiple cell and tissue types by a new affinity capture method
Description	Centrosomes are small cytoplasmic organelles that play fundamental roles in a range of cellular processes. Nearly all vertebrate cell types contain centrosomes, but whether centrosome composition and function differ between cell types, tissues and pathologies remains an outstanding question. Until now, probing centrosome composition has relied on proteomic profiling of centrosomes obtained by consecutive sucrose density gradient centrifugations, requiring up to one billion cells, thus rendering studies with multiple conditions cumbersome. Here we describe centrosome affinity capture (CAPture)-mass spectrometry (MS), a method that achieves high-coverage proteomes from 20-30 million cells. Utilising a biotin-labelled peptide derived from the CCDC61 protein, CAPture isolates intact centrosomes in a manner dependent on Ninein, a conserved centrosome component. CAPture-MS performs well across several untransformed and primary cell lines, thereby enabling comparisons of high-resolution centrosome proteomes. Using distal appendage proteins as an example, we demonstrate the utility of CAPture-MS for dissecting hierarchical interactions within the centrosome. Overall CAPture-MS represents a powerful tool to unveil temporal, regulatory, cell type- and tissue-specific changes in centrosome proteomes in health and disease.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_07:59:32.067.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Evangelia Papachristou
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	TMT6plex-126 reporter+balance reagent acylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2023-02-21 02:45:18	ID requested
1	2023-10-18 03:59:11	announced
⏵ 2	2023-11-14 07:59:32	announced	2023-11-14: Updated project metadata.
3	2024-10-22 06:06:11	announced	2024-10-22: Updated project metadata.

Dataset with its publication pending

submitter keyword: proteomics,centrosomes, capture MS

Fanni Gergely
contact affiliation	1. CRUK Cambridge Institute, Li Ka Shing Centre, University of Cambridge, United Kingdom 2. Department of Biochemistry, University of Oxford, UK
contact email	fanni.gergely@bioch.ox.ac.uk
lab head
Evangelia Papachristou
contact affiliation	Senior Scientific Assistant
contact email	Eva.Papachristou@cruk.cam.ac.uk
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/2023/10/PXD040309

PRIDE project URI

• PRIDE
  1. PXD040309
     1. Label: PRIDE project
     2. Name: Proteomic profiling of centrosomes across multiple cell and tissue types by a new affinity capture method