CCeMMP Special Seminar – Prof. Dr. Misha Kudryashev – Dec 2024

Subtomogram averaging from cryo-electron tomograms is a powerful method to determine structures of macromolecules in their native state. Outstanding applications to protein lattices, coats and ribosomes provided unique insights into their functions and even revealed interactions with small molecules in situ. For other macromolecules, such as membrane proteins, which are present in tomograms in limited numbers, the throughput of data processing and the processing time are key bottlenecks in obtaining high-resolution reconstructions. This is particularly the case for membrane proteins that are typically present in tomograms in moderate amounts.

Dr. Kudryashev will introduce tools that were developed in his lab for in situ structural biology with a focus on a large ion channel, RyR1, which is a part of the excitation-contraction coupling in muscle. TomoBEAR is a workflow for processing tomographic data utilizing common cryo-EM tools and original code that allows transparent near-automated tomographic pre-processing, alignment, reconstruction and particle identification followed by structural analysis.

In the second part of the talk, he will show recent results on understanding the molecular architecture of synaptic vesicles. Neurons grown on EM grids and purified synaptic vesicles were imaged by cryo-electron tomography. Individual proteins important for the function of synaptic vesicles, could be identified using this technique.

Prof. Dr. Misha Kudryashev

Max Delbrück Center for Molecular Medicine in the Helmholtz Society, Berlin, Germany