An extrusion strategy for the FeMo cofactor from nitrogenase: Towards synthetic iron-sulfur proteins

Lisandra L. Martin, Lee C. West, Bei Wu

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Iron-sulfur clusters are ubiquitous in biological systems, facilitating functions such as electron transfer (rubredoxins, ferredoxins, rieske centres), isomerization (aconitase) and small molecule activation such as dinitrogen reduction (nitrogenases). Of global importance and recently particular interest, is the iron-sulfur-containing iron-molybdenum cofactor (FeMoco) cluster that achieves the biological reduction of dinitrogen under mild conditions. This biologically unique cluster has proved difficult to investigate due to its extreme air sensitivity and the instability of the cluster's structural integrity, outside the protective protein matrix. Here, we report a model iron-sulfur cluster (Roussins black salt (NH4)[Fe4S3(NO)7]) that has been used to achieve the first example of a metal cluster (guest) embedded within a pseudo-protein, cyclodextrin (host). The product formed is supramolecular, that is, it contained no covalent bonds and was stabilized by predominantly entropy effects. Formation of a 1:1 complex between the host and the guest was established for the iron-sulfur cluster with either seven- or eight-membered cyclodextrins (β- or γ-cyclodextrin). A range of techniques was used to characterize the new complexes in both the solid and solution states. Electrospray mass spectra indicated the presence of parent ions of the host-guest complexes and electrochemistry was also used to define the redox behavior of the complexes. The iron-sulfur clusters were significantly more stable in the presence of the host cyclodextrin, as revealed by a negative shift for the reduction potential for the host-guest product. Using the β-cyclodextrin as host, the reduction potential of the iron-sulfur cluster shifted more negative by 60 mV; the effect was even more dramatic for the larger γ-cyclodextrin where the reduction potential for the cluster was shifted by 90 mV more negative than the 'unbound' [Fe4S3(NO)7]- cluster. This is the first example of a metal cluster, stabilized as a supramolecular complex in a 'host' environment outside of a covalently bonded protein matrix. Creating such stable environments for metal cofactors or clusters that otherwise spontaneously degrade or are catalytically inactive outside the protein matrix could have enormous practical value. Specific implications for the development of extrusion methods for FeMoco from nitrogenase are enormous, with previously difficult, high-energy molecular transformations, such as dinitrogen to ammonia, now more realistically accessible.

Original languageEnglish
Pages (from-to)5676-5686
Number of pages11
JournalEuropean Journal of Biochemistry
Issue number22
Publication statusPublished - 3 Dec 2001
Externally publishedYes


  • FeMo cofactor
  • Host-guest complexes
  • Iron-sulfur clusters
  • Supra-molecular
  • Synthetic proteins

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