Regensburg 2025 – scientific programme

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BP: Fachverband Biologische Physik

BP 17: Poster Session II

BP 17.59: Poster

Tuesday, March 18, 2025, 18:00–20:30, P4

Investigation into the dynamic structure of heat shock proteins using electrospray ion beam deposition and cryo-electron microscopy (ESIBD+cryoEM) — •Noor Naseeb, Lukas Eriksson, Jingjin Fan, Justin Benesch, and Stephan Rauschenbach — University of Oxford, Oxford, United Kingdom

The heat shock protein (HSP) family encompasses a wide variety of polydisperse proteins that act as chaperones in the cell as a means of preventing aggregation and misfolding of proteins under different forms of cellular stress. Standard methods, like X-ray crystallography (XRC), Nuclear magnetic resonance (NMR), and cryogenic electron microscopy (cryo-EM), lack the ability to properly study the structures of such dynamic and diverse proteins. To address this, we use electrospray ion beam deposition (ESIBD) that couples native mass spectrometry (MS), a chemically selective sample preparation technique, with cryo-EM. The combination allows for high-resolution visualization of a specific protein assembly by cryo-EM in their near-native state. Here we show that the chemically selective sample preparation technique via ESIBD enables structure determination of these dynamically assembled HSPs can be performed to better assess their structure, and therefore, function.

Keywords: Heat shock proteins; Cryogenic electron microscopy; Native Mass Spectrometry; Electrospray ion beam deposition; Protein structure determination