Panagiotis L. Kastritis

Exploring cellular organization in native cell extracts with high-resolution cryo-EM

Panagiotis L. Kastritis1,2

1 Institute of Biochemistry and Biotechnology; Biozentrum; and Interdisciplinary Research Center HALOmem, Martin-Luther University Halle-Wittenberg, Halle (Saale), Germany;
2 Institute of Chemical Biology, National Hellenic Research Foundation, Athens, Greece

Advances in electron microscopy have provided unprecedented access to the structural characterization of large, flexible, and heterogeneous complexes. Until recently, cryo-electron microscopy (cryo-EM) has been applied to understand molecular organization in either highly purified, isolated biomolecules or in situ. An emerging field is developing, bridging the gap between the two approaches, studying molecular organization in native cellular fractions. Due to the biochemical nature of the cell extract, functional assays, mass spectrometry, and single-particle cryo-EM analysis, combined with artificial intelligence, can provide multi-scale structure-function understanding. Here, I will present structural insights into previously elusive megadalton protein complexes uncovered via integrative analysis of cellular fractions. Overall, our methods and data provide a framework for further understanding cellular organization by systematically mapping the structure of endogenous biomolecular assemblies and their interactions within cellular fractions.

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