PXD018789

PXD018789 is an original dataset announced via ProteomeXchange.

Title	Retargeting from CR3 to the LFA-1 receptor uncovers the adenylyl cyclase enzymetranslocating segment of Bordetella adenylate cyclase toxin
Description	The Bordetella adenylate cyclase toxinhemolysin (CyaA) and the α-hemolysin (HlyA) of Escherichia coli both belong to the family of cytolytic pore-forming Repeats in ToXin (RTX) cytotoxins. HlyA preferentially binds the αLβ2 integrin LFA-1 (CD11a/CD18) of leukocytes and can promiscuously bind and permeabilize also a variety of other cells. CyaA bears an Nterminal adenylyl cyclase enzyme (AC) domain linked to a pore-forming RTX cytolysin (Hly) moiety and binds the complement receptor 3 (CR3, αMβ2, CD11b/CD18 or Mac-1) of myeloid phagocytes, penetrates their plasma membrane and delivers into cytosol the AC enzyme. We constructed a set of CyaA/HlyA chimeras and show that the CyaC-acylated segment and the CR3-binding RTX domain of CyaA can be functionally replaced by the much shorter HlyCacylated and LFA-1-binding RTX moiety of HlyA. Instead of binding CR3, a CyaA1710/HlyA411-1024 chimera bound the LFA-1 receptor and effectively delivered the AC enzyme into Jurkat lymphoblastoma T cells. At high chimera concentrations (25 nM), the interaction with LFA-1 was not required for CyaA1-710/HlyA411-1024 binding to CHO cells. However, interaction with the LFA-1 receptor strongly enhanced the specific capacity of the bound CyaA1-710/HlyA411-1024 chimera to penetrate cells and deliver the AC enzyme into their cytosol. Hence, interaction of the acylated RTX moiety of HlyA with LFA-1 promoted a productive membrane interaction of the chimera. These results allow to delimit the residues 400 to 710 of CyaA as the 'AC translocon' sufficient for translocation of the AC enzyme polypeptide across the plasma membrane of target cells
HostingRepository	PRIDE
AnnounceDate	2020-05-11
AnnouncementXML	Submission_2020-05-17_23:51:38.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	David Jurnečka
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562; scientific name: Bordetella pertussis Tohama I; NCBI TaxID: 257313;
ModificationList	palmitoylated residue; myristoylated residue; monohydroxylated residue
Instrument	Bruker Daltonics solarix series

Revision	Datetime	Status	ChangeLog Entry
0	2020-04-24 03:31:59	ID requested
1	2020-05-10 22:46:42	announced
⏵ 2	2020-05-17 23:51:38	announced	2020-05-18: Updated publication reference for PubMed record(s): 32393579.

Masin J, Osickova A, Jurnecka D, Klimova N, Khaliq H, Sebo P, Osicka R, adenylate cyclase toxin. J Biol Chem, 295(28):9349-9365(2020) [pubmed]

submitter keyword: AC domain translocation, acylation, acyltransferase, fatty acyl, integrin, RTX toxin

Prof. Peter Sebo, PhD.
contact affiliation	Institute of Microbiology of the CAS, v. v. i.
contact email	sebo@biomed.cas.cz
lab head
David Jurnečka
contact affiliation	Institute of Microbiology, Czech Academy of Sciences
contact email	david.jurnecka@biomed.cas.cz
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2020/05/PXD018789

PRIDE project URI

• PRIDE
  1. PXD018789
     1. Label: PRIDE project
     2. Name: Retargeting from CR3 to the LFA-1 receptor uncovers the adenylyl cyclase enzymetranslocating segment of Bordetella adenylate cyclase toxin