Oxygen-Sensitive Metalloprotein Structure Determination by Cryo-Electron Microscopy

Metalloproteins are involved in key cell processes such as photosynthesis, respiration, and oxygen transport. However, the presence of transition metals (notably iron as a component of [Fe-S] clusters) often makes these proteins sensitive to oxygen-induced degradation. Consequently, their study usua...

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Bibliographic Details
Published in:Biomolecules (Basel, Switzerland) Switzerland), 2022-03, Vol.12 (3), p.441
Main Authors: Cherrier, Mickaël V, Vernède, Xavier, Fenel, Daphna, Martin, Lydie, Arragain, Benoit, Neumann, Emmanuelle, Fontecilla-Camps, Juan C, Schoehn, Guy, Nicolet, Yvain
Format: Article
Language:English
Subjects:
Anaerobic conditions
anaerobic environment
Apoferritin
Apoferritins
Biochemistry, Molecular Biology
Carbon monoxide dehydrogenase
Chemical reactions
cryo-electron microscopy
Cryoelectron Microscopy - methods
Crystallization
Crystallography
Crystallography, X-Ray
Electron microscopy
Ferredoxin
Heavy metals
iron-sulfur cluster
Life Sciences
Macromolecules
Metabolism
Metalloproteins
Metals
Microscopy
Nitrogen
Oxidoreductase
Oxygen
Photosynthesis
Proteins
Pyruvic acid
Structural Biology
Transition metals
X-ray crystallography
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Summary:Metalloproteins are involved in key cell processes such as photosynthesis, respiration, and oxygen transport. However, the presence of transition metals (notably iron as a component of [Fe-S] clusters) often makes these proteins sensitive to oxygen-induced degradation. Consequently, their study usually requires strict anaerobic conditions. Although X-ray crystallography has been the method of choice for solving macromolecular structures for many years, recently electron microscopy has also become an increasingly powerful structure-solving technique. We have used our previous experience with cryo-crystallography to develop a method to prepare cryo-EM grids in an anaerobic chamber and have applied it to solve the structures of apoferritin and the 3 [Fe S ]-containing pyruvate ferredoxin oxidoreductase (PFOR) at 2.40 Å and 2.90 Å resolution, respectively. The maps are of similar quality to the ones obtained under air, thereby validating our method as an improvement in the structural investigation of oxygen-sensitive metalloproteins by cryo-EM.
ISSN:2218-273X
2218-273X
DOI:10.3390/biom12030441