Proteins gain their functional structure in the process of protein folding. Most proteins begin to fold co-translationally on the ribosome while they emerge from its exit tunnel during protein biosynthesis. This is a fundamental process for maintaining cellular proteostasis, failure of which results in polypeptide misfolding and diseases such as Parkinson’s, Alzheimer’s or other proteinopathies. Despite its importance, it remains poorly understood as it is a significant challenge to obtain high-resolution structural data of co-translational protein folding (coTF) using structural biology methods alone; particularly the description of the folding pathways is missing. Consequently, the use of accurate and
efficient computational techniques, especially in combination with NMR and cryo-EM experimental data used as restraints, for reweighting or validation, is imperative for a detailed understanding of protein folding in the cell. In this presentation, I will describe an integrative structural biology approach to study co-translational protein folding, combining bioinformatics, all-atom and coarse-grained molecular dynamics (MD) simulations with structural restraints from various experimental data (NMR chemical shifts, RDCs and cryo-EM maps) and applied it to provide a high-resolution description of snapshots of the protein biosynthesis. I will present the characterisation of the dynamics and interactions of both folded and intrinsically disordered nascent chains on the wild-type bacterial ribosome and the rationally designed ribosome based on bioinformatics analysis. Finally, I will provide the first structural insights into the co-translational folding intermediate that were experimentally confirmed and are the first step to fully characterising the co-translational folding pathway.