PXD025459

PXD025459 is an original dataset announced via ProteomeXchange.

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

Revision	Datetime	Status	ChangeLog Entry
0	2021-04-19 00:18:43	ID requested
⏵ 1	2021-08-26 07:09:44	announced

Dataset with its publication pending

submitter keyword: BRAVO AssayMap

Fe(III)-IMAC

TiO2

automation

phosphopeptide enrichment

phosphoproteomics

quantification

Listeria monocytogenes

Bacillus subtilis

Streptococcus pyogenes

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

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PRIDE project URI

• PRIDE
  1. PXD025459
     1. Label: PRIDE project
     2. Name: Automated Phosphopeptide Enrichment for Gram Positive Bacteria