Regensburg 2025 – scientific programme

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BP: Fachverband Biologische Physik

BP 30: Protein Structure and Dynamics

BP 30.10: Talk

Thursday, March 20, 2025, 17:45–18:00, H46

Native Electrospray Ion Beam Deposition for Atomic-level Structure Analysis of Membrane Protein — •Jingjin Fan, Tim Esser, Clare De'Ath, Lukas Eriksson, Abdul Aziz Qureshi, Abraham Abraham, Laurence Seeley, Lindsay Baker, Carol Robinson, and Stephan Rauschenbach — The Kavli Institute for Nanoscience Discovery, University of Oxford, Oxford OX1 3QU, United Kingdom

Membrane proteins play vital roles in cellular physiology, but their structural analysis remains challenging due to heterogeneity, flexible conformations, and demanding native conditions. To address these challenges, we established electrospray ion beam deposition (ESIBD) to directly couple native mass spectrometry (MS) with cryogenic electron microscopy (cryo-EM) for studying membrane protein structures.

Standard membrane proteins, including aquaporin Z (AqpZ) and ammonium transporter B (AmtB), were selected as testing models. By optimizing surfactant, the ion transfer in vacuum and the embedding of the proteins after landing we successfully manipulated membrane protein particles and achieved soft-landing on grids, evidenced by high-resolution imaging in cryoEM.

Our results demonstrate that the membrane structures can be preserved even in the absence of visible micelle.This molecular-level structural analysis captured by ESIBD in vacuum provides new insights into the correct folding of membrane proteins and understanding fundamental questions in structural biology.

Keywords: single molecule imaging; membrane protein; structural biology