Bordetella pertussis, B. parapertussis and B. bronchiseptica are Gram-negative bacteria that colonize the respiratory tracts of mammals. B. pertussis has adapted exclusively to the human host and is known to be the causative agent of whooping cough (pertussis). Bordetella bacteria use the type III secretion system (T3SS) to establish persistent colonization of the trachea and to modulate the host immune response. They encode a highly conserved type III secretion effector protein, BteA, which is responsible for the induction of rapid, non-apoptotic death in a wide range of mammalian cells. BteA, the only type III effector that has been identified in Bordetella, shows no homology with any other type III effectors. The overall goal is to determine BteA structure and to elucidate its involvement in mechanisms leading to cell death. Our specific aims are:
1. To express, purify and determine the 3D structure of BteA from B. bronchiseptica.
2. To analyze BteA structure and determine its functional sites and to determine the ability of BteA to bind ligands.
3. To locate BteA interacting proteins in the eukaryotic host cell by using BteA as bait and thus determine i) protein function; ii) the structure of BteA-target molecule complexes.
Cutting-edge technologies will be used for structure determination and functional analysis of clean protein samples (X-ray crystallography, structure prediction and functional point mutations) and for the investigation of ligand binding and host cell interactions (pull-down assays, mass spectrometry and surface plasmon resonance).
The outcome will be new targets for antibiotic development and a better understanding of protein-protein interactions, cell targeting and cell death. The research will enhance EU competitiveness in structural biology and the life sciences and in the long term will contribute to public health (treatment and prevention of whooping cough).
Fields of science
- medical and health scienceshealth sciencespublic and environmental healthepidemics preventionimmunisation
- natural sciencesbiological sciencesmicrobiologybacteriology
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteinsproteomics
- natural scienceschemical sciencesanalytical chemistrymass spectrometry
- natural sciencesbiological sciencesmolecular biologystructural biology
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