Biophysical Society Thematic Meeting | Trieste 2024

Emerging Theoretical Approaches to Complement Single-Particle Cryo-EM

Poster Abstracts

24-POS Board 24 PREDICTING PROBLEMS CAUSED BY THE AIR-WATER INTERFACE IN CRYO ELECTRON MICROSCOPY: INSIGHTS FROM MOLECULAR DYNAMICS SIMULATIONS

Abhilash Sahoo 1 ; Miro Astore 1 ; Sonya Hanson 1 ; Brato Chakrabarti 2 ; 1 Flatiron Institute, Center for Computational Biology, New York, NY, USA 2 International Centre for Theoretical Sciences, Bangalore, India

Cryo-electron microscopy (cryo-EM) has revolutionized structural biology and biophysics by enabling high-resolution analysis of diverse biomolecular structures. However, a persistent challenge in single particle cryo-EM is the phenomenon of preferred particle orientation, which limits the accuracy of three-dimensional structural reconstructions and hinders interpretation. This issue is often exacerbated by the interaction of proteins with the air-water interface (AWI) during specimen preparation. In this study, we employ coarse-grained molecular dynamics simulations to investigate this complex interplay between proteins, detergents, and the AWI. Specifically, we focus on the preferential partitioning and orientation behavior of proteins to the AWI and the influence of detergents with specific biophysical properties on this process. By mimicking the conditions encountered during cryo-EM sample preparation, we shed light on the mechanisms governing preferred orientations and partitioning at the AWI. This work advances our fundamental biophysical understanding of protein interactions at the AWI, and also offers practical implications for improving cryo-EM sample preparation protocols.

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