9CQZ
Azotobacter vinelandii Reduced MoFeP (C1 symmetry) obtained using the SPT Labtech chameleon of 20 mM sodium dithionite under Al's oil
Summary for 9CQZ
| Entry DOI | 10.2210/pdb9cqz/pdb |
| EMDB information | 45828 |
| Descriptor | Nitrogenase molybdenum-iron protein alpha chain, Nitrogenase molybdenum-iron protein beta chain, 3-HYDROXY-3-CARBOXY-ADIPIC ACID, ... (7 entities in total) |
| Functional Keywords | nitrogenase, femoco, nitrogen, p-cluster, metal binding protein |
| Biological source | Azotobacter vinelandii More |
| Total number of polymer chains | 4 |
| Total formula weight | 233239.17 |
| Authors | Cook, B.D.,Narehood, S.M.,McGuire, K.L.,Li, Y.,Tezcan, F.A.,Herzik, M.A. (deposition date: 2024-07-19, release date: 2025-04-30) |
| Primary citation | Cook, B.D.,Narehood, S.M.,McGuire, K.L.,Li, Y.,Akif Tezcan, F.,Herzik Jr., M.A. Preparation of oxygen-sensitive proteins for high-resolution cryoEM structure determination using blot-free vitrification. Nat Commun, 16:3528-3528, 2025 Cited by PubMed Abstract: High-quality grid preparation for single-particle cryogenic electron microscopy (cryoEM) remains a bottleneck for routinely obtaining high-resolution structures. The issues that arise from traditional grid preparation workflows are particularly exacerbated for oxygen-sensitive proteins, including metalloproteins, whereby oxygen-induced damage and alteration of oxidation states can result in protein inactivation, denaturation, and/or aggregation. Indeed, 99% of the current structures in the EMBD were prepared aerobically and limited successes for anaerobic cryoEM grid preparation exist. Current practices for anaerobic grid preparation involve a vitrification device located in an anoxic chamber, which presents significant challenges including temperature and humidity control, optimization of freezing conditions, costs for purchase and operation, as well as accessibility. Here, we present a streamlined approach that allows for the vitrification of oxygen-sensitive proteins in reduced states using an automated blot-free grid vitrification device - the SPT Labtech chameleon. This robust workflow allows for high-resolution structure determination of dynamic, oxygen-sensitive proteins, of varying complexity and molecular weight. PubMed: 40229244DOI: 10.1038/s41467-025-58243-1 PDB entries with the same primary citation |
| Experimental method | ELECTRON MICROSCOPY (2.19 Å) |
Structure validation
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