PXD022078-1

PXD022078 is an original dataset announced via ProteomeXchange.

Title	MS-based in situ proteomics reveals AMPylation of host proteins during bacterial infection
Description	Bacteria utilize versatile strategies to propagate infections within human cells, e.g. by the injection of effector proteins which alter crucial signaling pathways. One class of such virulence-associated proteins is involved in the AMPylation of eukaryotic Rho GTPases with devastating effects on viability. In order to get an inventory of AMPylated proteins, several technologies have been developed. However, as they were designed for the analysis of cell lysates, knowledge about AMPylation targets in living cells is largely lacking. We here implement a chemical-proteomic method for deciphering AMPylated host proteins in situ during bacterial infection. HeLa cells treated with a previously established cell permeable pronucleotide probe (pro-N6pA) were infected with Vibrio parahaemolyticus and modified host proteins were identified upon probe enrichment and LC-MS/MS analysis. Three already known targets of the AMPylator VopS - Rac1, RhoA and Cdc42 - could be confirmed and several other Rho GTPases were additionally identified. Applying an inactive VopS mutant validated these hits. The utility was further demonstrated by connecting the virulence phenotype of cell rounding with VopS mediated AMPylation of essential targets as well as deciphering the sites of modification. Overall, the methodology provides a reliable detection of host AMPylation in situ and thus a versatile tool in monitoring infection processes.
HostingRepository	PRIDE
AnnounceDate	2020-12-08
AnnouncementXML	Submission_2020-12-07_22:48:34.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Theresa Rauh
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	iodoacetamide derivatized residue
Instrument	Q Exactive; Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2020-10-19 08:30:32	ID requested
⏵ 1	2020-12-07 22:48:35	announced

Rauh T, Brameyer S, Kielkowski P, Jung K, Sieber SA, Proteomics Reveals AMPylation of Host Proteins during Bacterial Infection. ACS Infect Dis, 6(12):3277-3289(2020) [pubmed]

submitter keyword: Human, HeLa, LC-MS/MS, Infection, V. parahaemolyticus

Stephan Axel Sieber
contact affiliation	Department of Chemistry, Chair of Organic Chemistry II, Technical University of Munich, Germany
contact email	stephan.sieber@tum.de
lab head
Theresa Rauh
contact affiliation	Department of Chemistry, Technical University of Munich, Lichtenbergstraße 4,85748 Garching, Germany Chair of Organic Chemistry II
contact email	theresa.rauh@tum.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD022078
     1. Label: PRIDE project
     2. Name: MS-based in situ proteomics reveals AMPylation of host proteins during bacterial infection