A post-doc position is available in our laboratory of Molecular Parasitology to determine the structure of key proteins involved in the development of human Sleeping sickness by CryoEM. Experience in biochemistry is essential and in electron microscopy desirable. Please contact david.perez-morga@ulb.be for details.
Projet summary :
CryoEM structure of key proteins for the evolution of African trypanosomiasis.
Recently we have implemented CryoEM in our institute (IBMM-ULB) to allow structural characterization of proteins of biomedical relevance. We propose to use CryoEM to solve the structure of APOL1, SRA and TgSGP, key proteins involved in the arms race between trypanosomes and humans during the development of sleeping sickness. APOL1 is a human serum protein that kills non-pathogenic African trypanosomes, while SRA and TgSGP are proteins that protect pathogenic trypanosomes against the lethal effects of APOL1. We aim to understand how SRA and TgSGP neutralise the effects of APOL1 by functional and protein structural analyses.
Until recently, the method of choice for solving protein structure at atomic resolution was X-ray crystallography, however, its major challenge remains the generation of protein crystals that can properly diffract X-rays; a particularly difficult task with our target hydrophobic proteins. Recent technological developments such as dramatic improvements in detector technologies and in analysis software, have brought CryoEM to resolve the structure of proteins to atomic resolution (current record 1.2 Å at 300keV, 1.8 Å at 200keV). All the workflow and benchmarking validation, from sample preparation to generation of 3D maps and models, have been established in our lab; from single particle analysis (SPA) to electron-tomography (ET) at room temperature and cryo conditions (CryoET).
We propose to analyse by CryoET the interaction of APOL1 and TgSGP with phospholipid membranes, and to determine the protein structure of APOL, SRA and TgSGP by SPA.
Main reading :
Mechanism of Trypanosoma brucei gambiense resistance to human serum. Uzureau P, Uzureau S, Lecordier L, Fontaine F, Tebabi P, Homblé F, Grélard A, Zhendre V, Nolan DP, Lins L, Crowet JM, Pays A, Felu C, Poelvoorde P, Vanhollebeke B, Moestrup SK, Lyngsø J, Pedersen JS, Mottram JC, Dufourc EJ, Pérez-Morga D, Pays E. Nature. 2013 ; 501(7467) : 430-4. doi: 10.1038/nature12516.
The molecular arms race between African trypanosomes and humans. Pays E, Vanhollebeke B, Uzureau P, Lecordier L, Pérez-Morga D. Nature Rev Microbiol. 2014 ; 12(8):575-84. doi: 10.1038/nrmicro3298.