Spectroscopic Investigations of [FeFe] Hydrogenase Maturated with [(57)Fe2(adt)(CN)2(CO)4](2-)

J Am Chem Soc. 2015 Jul 22;137(28):8998-9005. doi: 10.1021/jacs.5b03270. Epub 2015 Jul 9.

Abstract

The preparation and spectroscopic characterization of a CO-inhibited [FeFe] hydrogenase with a selectively (57)Fe-labeled binuclear subsite is described. The precursor [(57)Fe2(adt)(CN)2(CO)4](2-) was synthesized from the (57)Fe metal, S8, CO, (NEt4)CN, NH4Cl, and CH2O. (Et4N)2[(57)Fe2(adt)(CN)2(CO)4] was then used for the maturation of the [FeFe] hydrogenase HydA1 from Chlamydomonas reinhardtii, to yield the enzyme selectively labeled at the [2Fe]H subcluster. Complementary (57)Fe enrichment of the [4Fe-4S]H cluster was realized by reconstitution with (57)FeCl3 and Na2S. The Hox-CO state of [2(57)Fe]H and [4(57)Fe-4S]H HydA1 was characterized by Mössbauer, HYSCORE, ENDOR, and nuclear resonance vibrational spectroscopy.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Carbon Monoxide / metabolism
  • Catalytic Domain
  • Chlamydomonas reinhardtii / chemistry
  • Chlamydomonas reinhardtii / enzymology*
  • Chlamydomonas reinhardtii / metabolism
  • Electron Spin Resonance Spectroscopy*
  • Hydrogenase / antagonists & inhibitors
  • Hydrogenase / chemistry*
  • Hydrogenase / metabolism
  • Iron Compounds / chemistry*
  • Iron Isotopes / chemistry
  • Iron-Sulfur Proteins / antagonists & inhibitors
  • Iron-Sulfur Proteins / chemistry*
  • Iron-Sulfur Proteins / metabolism
  • Models, Molecular
  • Spectroscopy, Mossbauer*

Substances

  • Iron Compounds
  • Iron Isotopes
  • Iron-Sulfur Proteins
  • Carbon Monoxide
  • iron hydrogenase
  • Hydrogenase