Document Type
Article
Author ORCID Identifier
Bruno Cuevas-Zuviría https://orcid.org/0000-0003-1479-9442
Franka Detemple https://orcid.org/0009-0000-0395-7527
Kaustubh Amritkar https://orcid.org/0009-0000-5270-3451
Lance C. Seefeldt https://orcid.org/0000-0002-6457-9504
Betül Kaçar https://orcid.org/0000-0002-0482-2357
Journal/Book Title
eLife
Publication Date
9-11-2025
Publisher
eLife Sciences Publications Ltd.
Journal Article Version
Version of Record
Volume
14
First Page
1
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.
Last Page
22
Abstract
Previously, we identified the only dinitrogen reduction mechanism known to date as an ancient feature conserved from nitrogenase ancestors, which we directly tested by resurrecting and integrating synthetic ancestral nitrogenases into the genome of Azotobacter vinelandii (Garcia et al., 2023), a genetically tractable, nitrogen-fixing model bacterium. Here, we extend this paleomolecular approach to investigate the structural evolution of nitrogenase over billions of years of evolution by combining phylogenetics, ancestral sequence reconstruction, protein crystallography, and deep-learning based predictions. This study reveals that nitrogenase, while maintaining a conserved multimeric core, evolved novel modular features aligned with major environmental transitions, suggesting that subtle distal changes and transient regulatory adaptations were key to its long-term persistence and to shaping protein evolution over geologic time. The framework established here provides a foundation for identifying structural constraints that governed ancient proteins and for situating their sequences and structures within phylogenetic and environmental contexts across time.
Recommended Citation
Bruno Cuevas Zuviría, Franka Detemple, Kaustubh Amritkar, Amanda K Garcia, Lance Seefeldt, Oliver Einsle, Betül Kaçar (2025) Structural evolution of nitrogenase over 3 billion years eLife 14:RP105613. https://doi.org/10.7554/eLife.105613.4