Integrative structural biology of cell extracts
Eymeline Pageot, Maria Bacia, Irina Gutsche, Ambroise Desfosses
This innovative approach bridges the gap between the high resolution structural characterisation of highly purified, isolated biomolecules and in situ electron tomography. By removing the need for overexpression and purification, structural biology of native cell extracts holds unparalleled potential for applications. Emerging pathogens with unannotated genomes, uncultivable organisms, environmental samples containing numerous organisms—this method relies on cryo-EM imaging of heterogeneous fractions containing hundreds of proteins and complexes. It aims to simultaneously solve a large number of high resolution structures, annotate the corresponding genomes with the sequences derived thereof, and identify new essential complexes in organisms that are absent from databases. The challenge is the throughput and automation of this process : currently, it takes months to process a single heterogeneous dataset of this type, whereas it would be necessary to combine dozens of datasets corresponding to various molecular weight ranges to create a sufficiently complete atlas of the endogenous structures of a target organism. This project aims to lay the foundations for automation by exploring each aspect : from optimizing image acquisition to automating image processing for 3D reconstruction, and implementing AI-based tools for the identification and atomic interpretation of the solved structures.
Collaborations :
– Yohann Couté, EdyP, IRIG, Grenoble
– Linda Sandblad and UCEM, Umeå University, Sweden
– Virgile Adam, I2SR, IBS