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Title | Oxygen-Sensitive Metalloprotein Structure Determination by Cryo-Electron Microscopy. |
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Journal, issue, pages | Biomolecules, Vol. 12, Issue 3, Year 2022 |
Publish date | Mar 12, 2022 |
Authors | Mickaël V Cherrier / Xavier Vernède / Daphna Fenel / Lydie Martin / Benoit Arragain / Emmanuelle Neumann / Juan C Fontecilla-Camps / Guy Schoehn / Yvain Nicolet / |
PubMed Abstract | 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] ...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 [FeS]-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. |
External links | Biomolecules / PubMed:35327633 / PubMed Central |
Methods | EM (single particle) |
Resolution | 2.4 - 2.9 Å |
Structure data | EMDB-13487: Horse spleen apoferritin frozen under anaerobic conditions EMDB-13493, PDB-7plm: |
Chemicals | ChemComp-SF4: ChemComp-TPP: ChemComp-CA: ChemComp-MG: ChemComp-HOH: |
Source |
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Keywords | OXIDOREDUCTASE / 4Fe-4S / anaerobic / PYRUVATE CATABOLISM / Electron Transport |