PXD025459 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Automated Phosphopeptide Enrichment for Gram Positive Bacteria |
Description | Protein phosphorylation in prokaryotes has gained more attention in recent years as several studies linked it to regulatory and signalling functions indicating an importance similar to protein phosphorylation in eukaryotes. Studies on bacterial phosphorylation have so far been conducted using manual or HPLC-supported phosphopeptide enrichment while automation of phosphopeptide enrichment has been established in eukaryotes, allowing for high throughput sampling. To facilitate the prospect of studying bacterial phosphorylation on a systems‐level we here establish an automated Ser/Thr/Tyr phosphopeptide enrichment workflow on the Agilent AssayMap platform. We present optimized buffer conditions for TiO2 and Fe(III)NTA-IMAC based enrichment, and the most advantageous, species specific starting amount for S. pyogenes, L. monocytogenes, and B. subtilis. Our data represents, to the best of our knowledge, the largest phosphoproteome identified by a single study for each of these bacteria. For higher sample amounts (> 250µg) we observed a superior performance of Fe(III)NTA cartridges, while more peptides were identified from smaller sample amounts using TiO2-based enrichment. Both cartridges largely enrich the same set of phosphopeptides suggesting no improvement of peptide yield from complementary use of both cartridges. Distribution of S/T/Y phosphorylation varied between bacterial strains with threonine being the main site of phosphorylation in S. pyogenes, while in B.subtilis and L. monocytogenes showed the highest percentage of phosphorylation at serine. |
HostingRepository | PRIDE |
AnnounceDate | 2021-08-26 |
AnnouncementXML | Submission_2021-08-26_07:09:43.898.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Christian Frese |
SpeciesList | scientific name: Listeria monocytogenes EGD; NCBI TaxID: 1334565; |
ModificationList | monohydroxylated residue; iodoacetamide derivatized residue |
Instrument | Orbitrap Fusion Lumos; Orbitrap Exploris 480 |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2021-04-19 00:18:43 | ID requested | |
⏵ 1 | 2021-08-26 07:09:44 | announced | |
Publication List
Dataset with its publication pending |
Keyword List
submitter keyword: BRAVO AssayMap |
Fe(III)-IMAC |
TiO2 |
automation |
phosphopeptide enrichment |
phosphoproteomics |
quantification |
Listeria monocytogenes |
Bacillus subtilis |
Streptococcus pyogenes |
Contact List
Christian Karl Frese |
contact affiliation | Max Planck Unit for the Science of Pathogens, Chariteplatz 1, 10117 Berlin, Germany |
contact email | frese@mpusp.mpg.de |
lab head | |
Christian Frese |
contact affiliation | Max Planck Unit for the Science of Pathogens |
contact email | frese@mpusp.mpg.de |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD025459
- Label: PRIDE project
- Name: Automated Phosphopeptide Enrichment for Gram Positive Bacteria