Article thumbnail

Cell-free H-cluster Synthesis and [FeFe] Hydrogenase Activation: All Five CO and CN− Ligands Derive from Tyrosine

By Jon M. Kuchenreuther, Simon J. George, Celestine S. Grady-Smith, Stephen P. Cramer and James R. Swartz

Abstract

[FeFe] hydrogenases are promising catalysts for producing hydrogen as a sustainable fuel and chemical feedstock, and they also serve as paradigms for biomimetic hydrogen-evolving compounds. Hydrogen formation is catalyzed by the H-cluster, a unique iron-based cofactor requiring three carbon monoxide (CO) and two cyanide (CN−) ligands as well as a dithiolate bridge. Three accessory proteins (HydE, HydF, and HydG) are presumably responsible for assembling and installing the H-cluster, yet their precise roles and the biosynthetic pathway have yet to be fully defined. In this report, we describe effective cell-free methods for investigating H-cluster synthesis and [FeFe] hydrogenase activation. Combining isotopic labeling with FTIR spectroscopy, we conclusively show that each of the CO and CN− ligands derive respectively from the carboxylate and amino substituents of tyrosine. Such in vitro systems with reconstituted pathways comprise a versatile approach for studying biosynthetic mechanisms, and this work marks a significant step towards an understanding of both the protein-protein interactions and complex reactions required for H-cluster assembly and hydrogenase maturation

Topics: Research Article
Publisher: Public Library of Science
OAI identifier: oai:pubmedcentral.nih.gov:3105041
Provided by: PubMed Central

To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.

Suggested articles

Citations

  1. (2010). [FeFe]-Hydrogenase Cyanide Ligands Derived From S-AdenosylmethionineDependent Cleavage of Tyrosine.
  2. (2010). A glycyl free radical as the precursor in the synthesis of carbon monoxide and cyanide by the [FeFe]-hydrogenase maturase HydG.
  3. (2006). A radical solution for the biosynthesis of the H-cluster of hydrogenase.
  4. (2005). Biochemical characterization of the HydE and HydG iron-only hydrogenase maturation enzymes from Thermatoga maritima.
  5. (2008). Cell-free synthesis and maturation of [FeFe] hydrogenases.
  6. (2008). Deletion of iscR stimulates recombinant clostridial Fe-Fe hydrogenase activity and H2-accumulation in Escherichia coli BL21(DE3).
  7. (2002). Direct comparison of the electrocatalytic oxidation of hydrogen by an enzyme and a platinum catalyst. Chem Commun (Camb).
  8. (2004). Discovery of two novel radical S-adenosylmethionine proteins required for the assembly of an active [Fe] hydrogenase.
  9. (2008). Dithiomethylether as a ligand in the hydrogenase h-cluster.
  10. (1996). Escherichia coli contains a protein that is homologous in function and N-terminal sequence to the protein encoded by the nifS gene of Azotobacter vinelandii and that can participate in the synthesis of the Fe-S cluster of dihydroxy-acid dehydratase.
  11. (2007). Evidence for nifU and nifS participation in the biosynthesis of the iron-molybdenum cofactor of nitrogenase.
  12. (2009). From hydrogenases to noble metal-free catalytic nanomaterials for H2 production and uptake.
  13. (2010). High-yield expression of heterologous [FeFe] hydrogenases in Escherichia coli.
  14. (2007). Highyield hydrogen production from starch and water by a synthetic enzymatic pathway.
  15. (2008). HydF as a scaffold protein in [FeFe] hydrogenase H-cluster biosynthesis.
  16. (2007). In vitro activation of [FeFe] hydrogenase: new insights into hydrogenase maturation.
  17. (2007). In vitro synthesis of the iron-molybdenum cofactor of nitrogenase from iron, sulfur, molybdenum, and homocitrate using purified proteins.
  18. (2002). Infrared studies of the CO-inhibited form of the Fe-only hydrogenase from Clostridium pasteurianum I: examination of its light sensitivity at cryogenic temperatures.
  19. (2006). NifB-dependent in vitro synthesis of the iron-molybdenum cofactor of nitrogenase.
  20. (2007). Occurrence, classification, and biological function of hydrogenases: an overview.
  21. (2010). Quantum refinement of [FeFe] hydrogenase indicates a dithiomethylamine ligand.
  22. (2009). Selforganized photosynthetic nanoparticle for cell-free hydrogen production.
  23. (2010). Stepwise [FeFe]-hydrogenase H-cluster assembly revealed in the structure of HydA(DeltaEFG).
  24. (2009). Structural and functional analogues of the active sites of the [Fe]-, [NiFe]-, and [FeFe]-hydrogenases.
  25. (2003). Taming of a poison: biosynthesis of the NiFe-hydrogenase cyanide ligands.
  26. (2006). The [Fe-Fe]-hydrogenase maturation protein HydF from Thermotoga maritima is a GTPase with an iron-sulfur cluster.
  27. (2010). The [FeFe]-hydrogenase maturase HydF from Clostridium acetobutylicum contains a CO and CNligated iron cofactor.
  28. (2011). The [FeFe]-hydrogenase maturation protein HydF contains a H-cluster like [4Fe4S]-2Fe site.
  29. (2006). The active site of the [FeFe]-hydrogenase from Desulfovibrio desulfuricans. II. Redox properties, light sensitivity and CO-ligand exchange as observed by infrared spectroscopy.
  30. (1998). The hydrogen hypothesis for the first eukaryote.
  31. (2009). The role of the maturase HydG in [FeFe]-hydrogenase active site synthesis and assembly.
  32. (2007). Thiamine biosynthesis in Escherichia coli: identification of the intermediate and byproduct derived from tyrosine.
  33. (2004). Thiamine biosynthesis in Escherichia coli: in vitro reconstitution of the thiazole synthase activity.
  34. (2009). Tyrosine, Cysteine, and SAdenosyl Methionine Stimulate In Vitro [FeFe] Hydrogenase Activation.
  35. (2009). Volbeda A