PXD004681

PXD004681 is an original dataset announced via ProteomeXchange.

Title	Probing the global kinome and phosphoproteome in Chlamydomonas reinhardtii via sequential enrichment and quantitative proteomics
Description	Identifying dynamic protein phosphorylation events is critical for understanding kinase/phosphatase regulated signaling pathways. To date, protein phosphorylation and kinase expression have been examined independently in photosynthetic organisms. Presented here is a method to study the global kinome and phosphoproteome in tandem for the first time in a photosynthetic organism, the model alga Chlamydomonas reinhardtii (Chlamydomonas), using mass spectrometry-based label-free proteomics. A dual enrichment strategy first targets intact protein kinases via capture on immobilized multiplexed inhibitor beads (MIBs) with subsequent proteolytic digestion of unbound proteins and peptide-based phosphorylation enrichment. To increase depth of coverage, both data-dependent and data-independent (via SWATH) mass spectrometric acquisitions were performed to obtain a >50% increase in coverage of the enriched Chlamydomonas kinome over coverage found with no enrichment. The quantitative phosphoproteomic dataset yielded 2,304 phosphopeptides and 1,765 localized phosphosites with excellent reproducibility across biological replicates (90% of quantified sites with coefficient of variation below 11%). This approach enables simultaneous investigation of kinases and phosphorylation events at the global level to facilitate understanding of kinase networks and their influence in cell signaling events.
HostingRepository	PRIDE
AnnounceDate	2016-10-05
AnnouncementXML	Submission_2016-10-05_06:05:49.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD004681
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Evan McConnell
SpeciesList	scientific name: Chlamydomonas reinhardtii; NCBI TaxID: 3055;
ModificationList	Phospho; Deamidated; Oxidation; Acetyl; Carbamidomethyl
Instrument	TripleTOF 5600

Revision	Datetime	Status	ChangeLog Entry
0	2016-07-29 07:26:18	ID requested
⏵ 1	2016-10-05 06:05:51	announced

Werth EG, McConnell EW, Gilbert TS, Couso Lianez I, Perez CA, Manley CK, Graves LM, Umen JG, Hicks LM, Probing the global kinome and phosphoproteome in Chlamydomonas reinhardtii via sequential enrichment and quantitative proteomics. Plant J, 89(2):416-426(2017) [pubmed]

curator keyword: Biological

submitter keyword: Mass spectrometry, Quantitative label-free proteomics, Kinome, Phosphoproteomics, Chlamydomonas reinhardtii

Leslie M. Hicks
contact affiliation	Department of Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599 USA
contact email	lmhicks@email.unc.edu
lab head
Evan McConnell
contact affiliation	Graduate Research Assistant
contact email	evanmcc@live.unc.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD004681
     1. Label: PRIDE project
     2. Name: Probing the global kinome and phosphoproteome in Chlamydomonas reinhardtii via sequential enrichment and quantitative proteomics