MicroED structure of lipid-embedded mammalian mitochondrial voltage-dependent anion channel

• DOI: 10.1073/pnas.2020010117
• Publication Date: 2020-12-08T17:15:26Z
• Authors: Michael W. Martyn...; Farha Khan; Johan Hattne; Jeff Abramson; Tamir Gonen
• Keywords: 0301 basic medicine; Microscopy; Protein Conformation; 1.1 Normal biological development and functioning; Cryoelectron Microscopy; Electron; Lipids; cryoEM; Mitochondrial Proteins; Mice; 03 medical and health sciences; FIB/SEM; Microscopy, Electron, Transmission; Underpinning research; microcrystal electron diffraction; Microscopy, Electron, Scanning; Transmission; Animals; Voltage-Dependent Anion Channels; Scanning; Crystallization; MicroED; bicelle crystallization

Significance Microcrystal electron diffraction (MicroED) is an electron cryo-microscopy (cryoEM) method for determining structures using submicron crystals. Until now, determining structures of membrane proteins by MicroED required that the protein crystals be in a solution amenable to standard cryoEM blotting and vitrification protocols. Here, we show that membrane protein microcrystals grown in a viscous bicelle mixture can become amenable to MicroED analyses by using modified blotting procedures combined with focused ion-beam milling. Our findings provide a basis for solving membrane protein structures using crystals embedded in a viscous media by MicroED.