PXD029827

PXD029827 is an original dataset announced via ProteomeXchange.

Title	In-depth characterization of the Clostridioides difficile phosphoproteome to identify Ser/Thr kinases substrates
Description	Clostridioides difficile is the leading cause of intestinal nosocomial post-antibiotic infections in adults. During infection, the bacterium must rapidly respond and adapt to the host environment by using survival strategies. Protein phosphorylation is a reversible post-translational modification employed ubiquitously for signal transduction and cellular regulation. Recently, Hanks-type Serine/Threonine kinases (STKs) and phosphatases (STPs), have emerged as important players in bacterial cell signaling and pathogenicity. However, characterization of STKs targets in prokaryotes remained a difficult challenge. Here, we applied and adapted the TiO2 phosphopeptide enrichment approach to determine the phosphoproteome of C. difficile. We have identified and quantified 2497 proteins representing 63,1% of the theoretical proteome. Among these proteins, 649 contained phosphorylated peptides with a localization probability >0,75. This pathogen encodes two STKs (PrkC and CD2148) and one associated phosphatase (STP), PrkC was shown to be crucial in the regulation of cell wall homeostasis, antibiotic resistance and cell division. Comparative phosphoproteomics of wild-type, prkC, CD2148, stp and double prkC CD2148 strains were performed in order to determinate STKs targets. 104 proteins were identified as specific substrates of PrkC, 46 of CD2148 and 94 of both kinases. Using a combination of in vivo and in vitro approaches, FtsK and Spo0A were validated as a direct target of PrkC. This study provides a detailed mapping of kinase-substrate relationships which may aid in the identification of new biomarkers and therapeutic targets.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_08:30:27.077.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Thibaut Douché
SpeciesList	scientific name: Peptoclostridium difficile (strain 630) (Clostridium difficile); NCBI TaxID: 272563;
ModificationList	L-methionine (S)-sulfoxide; phosphorylated residue; N-acylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2021-11-20 08:35:54	ID requested
1	2022-12-08 05:14:48	announced
⏵ 2	2023-11-14 08:30:28	announced	2023-11-14: Updated project metadata.

Garcia-Garcia T, Douch, é T, Giai Gianetto Q, Poncet S, El Omrani N, Smits WK, Cuenot E, Matondo M, Martin-Verstraete I, In-Depth Characterization of the Clostridioides difficile Phosphoproteome to Identify Ser/Thr Kinase Substrates. Mol Cell Proteomics, 21(11):100428(2022) [pubmed]

submitter keyword: Phosphoproteomics, QExactive HF,Clostridium difficile, Kinase

Mariette Matondo
contact affiliation	Institut Pasteur, Université de Paris, CNRS USR2000, Proteomics Platform, Mass Spectrometry for Biology Unit, 75015 Paris, France
contact email	mariette.matondo@pasteur.fr
lab head
Thibaut Douché
contact affiliation	Institut Pasteur
contact email	thibaut.douche@pasteur.fr
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD029827
     1. Label: PRIDE project
     2. Name: In-depth characterization of the Clostridioides difficile phosphoproteome to identify Ser/Thr kinases substrates