Oxygen-sensitive metalloprotein structure determination by cryo-electron microscopy - Archive ouverte HAL Access content directly
Journal Articles Biomolecules Year : 2022

Oxygen-sensitive metalloprotein structure determination by cryo-electron microscopy

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] 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$_4$S$_4$]-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.
Fichier principal
Vignette du fichier
cherr1.pdf (5.85 Mo) Télécharger le fichier
Origin : Publisher files allowed on an open archive

Dates and versions

hal-03615151 , version 1 (23-03-2022)

Identifiers

Cite

Mickaël Cherrier, Xavier Vernède, Daphna Fenel, Lydie Martin, Benoit Arragain, et al.. Oxygen-sensitive metalloprotein structure determination by cryo-electron microscopy. Biomolecules, 2022, 12 (3), pp.441. ⟨10.3390/biom12030441⟩. ⟨hal-03615151⟩
81 View
76 Download

Altmetric

Share

Gmail Facebook Twitter LinkedIn More