Trapping H- bound to the nitrogenase FeMo-cofactor active site during H2 evolution: Characterization by ENDOR spectroscopy

Robert Y. Igarashi, Mikhail Laryukhin, Patricia C. Dos Santos, Hong In Lee, Dennis R. Dean*, Lance C. Seefeldt, Brian M. Hoffman

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

167 Scopus citations


We here show that the iron-molybdenum (FeMo)-cofactor of the nitrogenase α-70Ile molybdenum-iron (MoFe) protein variant accumulates a novel S = 1/2 state that can be trapped during the reduction of protons to H2. 1,2H-ENDOR measurements disclose the presence of two protons/hydrides (H+/-) whose hypertine tensors have been determined from two-dimensional field-frequency 1H ENDOR plots. The two H +/- have large isotropic hyperfine couplings, Aiso ≈ 23 MHz, which shows they are bound to the cofactor. The favored analysis for these plots indicates that the two H+/- have the same principal values, which indicates that they are chemically equivalent. The tensors are further related to each other by a permutation of the tensor components, which indicates an underlying symmetry of binding relative to the cofactor. At present, no model for the structure of the iron-molybdenum (FeMo)-cofactor in the S = 1/2 state trapped during the reduction of H+ can be shown unequivocally to satisfy all of the constraints generated by the ENDOR analysis. The data disfavors any model that involves protonation of sulfides, and thus suggests that the intermediate instead contains two chemically equivalent bound hydrides; it appears unlikely that these are terminal monohydrides.

Original languageEnglish (US)
Pages (from-to)6231-6241
Number of pages11
JournalJournal of the American Chemical Society
Issue number17
StatePublished - May 4 2005

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry


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