PXD003113

PXD003113 is an original dataset announced via ProteomeXchange.

Title	Stoichiometry of the bacterial type III secretion export apparatus
Description	Bacterial type III secretion systems are cell envelope-spanning effector protein-delivery machines essential for colonization and survival of many Gram-negative pathogens and symbionts. The membrane-embedded core unit of these secretion systems is termed needle complex. The needle complex is composed of a base that anchors the machinery to the inner and outer membranes, a hollow filament formed by inner rod and needle subunits that serves as conduit for substrate proteins, and a membrane-embedded export apparatus facilitating substrate translocation. While the stoichiometry of the base and of the major export apparatus protein have been revealed by structural analyses, the stoichiometries of the other export apparatus components and of the inner rod remain unknown. We employed peptide concatenated standard and absolute quantification-based strategies to analyze the stoichiometry of the entire needle complex by mass spectrometry. Here we provide evidence that the export apparatus of the type III secretion system encoded on Salmonella pathogenicity island 1 contains 5 SpaP, 1 SpaQ, 1 SpaR, and 1 SpaS. We have corroborated the previously suggested stoichiometry of 9 InvA per needle complex and describe a loose association of InvA with other needle complex components that may reflect its function. These numbers indicate that the inner membrane patch of the needle complex base houses 104 transmembrane domains in total, a dense assembly whose function in the secretion process we merely understand. Furthermore, we present evidence that not more than 6 PrgJ form the inner rod of the needle complex.
HostingRepository	PRIDE
AnnounceDate	2016-02-25
AnnouncementXML	Submission_2016-02-25_01:55:44.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Nicolas Nalpas
SpeciesList	scientific name: Salmonella enterica subsp. enterica serovar Typhimurium str. SL1344; NCBI TaxID: 216597;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive; LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2015-10-27 08:27:31	ID requested
⏵ 1	2016-02-25 01:55:45	announced

Zilkenat S, Franz-Wachtel M, Stierhof YD, Gal, á, n JE, Macek B, Wagner S, Determination of the Stoichiometry of the Complete Bacterial Type III Secretion Needle Complex Using a Combined Quantitative Proteomic Approach. Mol Cell Proteomics, 15(5):1598-609(2016) [pubmed]

curator keyword: Biological

submitter keyword: type III secretion, mass spectrometry, stoichiometry, Salmonella

Samuel Wagner
contact affiliation	University of Tübingen Interfaculty Institute of Microbiology and Infection Medicine (IMIT) Section of Cellular and Molecular Microbiology Elfriede-Aulhorn-Str. 6 72076 Tübingen Germany
contact email	samuel.wagner@med.uni-tuebingen.de
lab head
Nicolas Nalpas
contact affiliation	Tuebingen University
contact email	nalpas.nicolas@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD003113
     1. Label: PRIDE project
     2. Name: Stoichiometry of the bacterial type III secretion export apparatus