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TitleStructural insights into metallocluster trafficking in the nitrogenase assembly scaffold NifEN.
Journal, issue, pagesNat Catal, Vol. 9, Issue 3, Page 281-294, Year 2026
Publish dateMar 3, 2026
AuthorsBryan Neumann / Kristal A Brandon / Robert Quechol / Diana S Suder / Chi Chung Lee / Yimo Yang / Kamil Górecki / Jared A Wiig / Yilin Hu / Shane Gonen / Markus W Ribbe /
PubMed AbstractNitrogenase catalyses small-molecule activation, and has great relevance to agronomy, environment and energy. Understanding the assembly of the complex nitrogenase cofactor is a decades-long goal in ...Nitrogenase catalyses small-molecule activation, and has great relevance to agronomy, environment and energy. Understanding the assembly of the complex nitrogenase cofactor is a decades-long goal in the field, and structural insights into this process remain scarce. Here we report a cryogenic electron microscopy (cryo-EM) study of heterologously expressed NifEN, a key player converting the precursor (L-cluster) to a mature cofactor (M-cluster). Structural analyses of apo- and holo-NifEN demonstrate major conformational changes triggered by L-cluster incorporation. Further examinations of NifEN structures with inwardly and outwardly bound L-clusters, coupled with supporting mutational studies, AlphaFold 3 predictions and negative-stain EM analyses of NifEN complexed with upstream (NifB) and downstream (NifH) assembly partners, reveal a tunnel linking NifEN with NifB and NifH, with NifEN serving as a dynamic hub that coordinates L-cluster reception, maturation and delivery via conformation-gated metallocluster trafficking.
External linksNat Catal / PubMed:41908675 / PubMed Central
MethodsEM (single particle)
Resolution14.8 - 19.0 Å
Structure data

EMDB-73406: Negative stained A. vinelandii NifEN-B' fusion
Method: EM (single particle) / Resolution: 19.0 Å

EMDB-73407: Negative stained A. vinelandii NifEN/NifH ADPxAIF4- stabilized complex
Method: EM (single particle) / Resolution: 14.8 Å

Source
  • Azotobacter vinelandii DJ (bacteria)

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