Cryo-EM for Structure Determination of Mitochondrial Ribosome Samples

Hauke S Hillen

doi:10.1007/978-1-0716-3171-3_6

Cryo-EM for Structure Determination of Mitochondrial Ribosome Samples

Methods Mol Biol. 2023:2661:89-100. doi: 10.1007/978-1-0716-3171-3_6.

Author

Hauke S Hillen^{1

2

3}

Affiliations

¹ Department of Cellular Biochemistry, University Medical Center Göttingen, Göttingen, Germany. hauke.hillen@med.uni-goettingen.de.
² Research Group Structure and Function of Molecular Machines, Max Planck Institute for Multidisciplinary Sciences, Göttingen, Germany. hauke.hillen@med.uni-goettingen.de.
³ Cluster of Excellence 'Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells' (MBExC), University of Göttingen, Göttingen, Germany. hauke.hillen@med.uni-goettingen.de.

PMID: 37166633
DOI: 10.1007/978-1-0716-3171-3_6

Abstract

Single-particle cryoelectron microscopy (cryo-EM) allows structure determination of large macromolecular complexes from conformationally and compositionally heterogeneous mixtures of particles. This technique has been used to reveal the architecture of the mitochondrial ribosome and to visualize transient states that occur during the translation cycle or during mitoribosome biogenesis. Here, we outline an exemplary workflow for the analysis of single-particle cryo-EM data of human mitoribosome samples. In addition, we provide an example dataset which can be used for training purposes alongside the protocol.

Keywords: Mitochondrial ribosome; image processing; sample preparation; single-particle cryo-EM.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cryoelectron Microscopy / methods
Humans
Image Processing, Computer-Assisted / methods
Macromolecular Substances / chemistry
Mitochondrial Ribosomes*
Single Molecule Imaging*

Substances

Macromolecular Substances