PXD014593

PXD014593 is an original dataset announced via ProteomeXchange.

Title	pFAIMS: An optimized workflow for quantitative multiplexed phosphorylation analysis using high-Field Asymmetric waveform Ion Mobility Spectrometry (FAIMS)
Description	Protein phosphorylation is vital for the regulation of cellular signaling. Isobaric tag-based proteomic techniques, such as tandem mass tags (TMT), can measure the relative phosphorylation states of peptides in a multiplexed format. However, the overall low stoichiometry of protein phosphorylation constrains the analytical depth of phosphopeptide analysis by mass spectrometry, thereby requiring robust and sensitive workflows. Here we evaluate and optimize high-Field Asymmetric waveform Ion Mobility Spectrometry (FAIMS) coupled to Orbitrap Tribrid mass spectrometers for the analysis of TMT10plex-labeled phosphopeptides. We determined that using FAIMS-SPS-MS3 with three compensation voltages (CV) in a single method minimizes inter-CV overlap and maximizes peptide coverage (e.g., CV=-40V/-60V/-80V) and that consecutive analyses using CID-MSA and HCD fragmentation at the MS2 stage increases the depth of phosphorylation analysis.
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_04:04:13.574.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Joao Paulo
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion Lumos; Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2019-07-12 08:14:00	ID requested
1	2020-01-10 05:54:21	announced
⏵ 2	2024-10-22 04:04:14	announced	2024-10-22: Updated project metadata.

Dataset with its publication pending

submitter keyword: FAIMS, TMT, Lumos, SL-TMT, isobaric,phosphorylation

Joao A. Paulo
contact affiliation	Joao A. Paulo Cell Biology Harvard Medical School Boston, MA 02115
contact email	joao_paulo@post.harvard.edu
lab head
Joao Paulo
contact affiliation	Harvard Medical School
contact email	joao_paulo@post.harvard.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD014593
     1. Label: PRIDE project
     2. Name: pFAIMS: An optimized workflow for quantitative multiplexed phosphorylation analysis using high-Field Asymmetric waveform Ion Mobility Spectrometry (FAIMS)