HydG, the “Dangler” Iron, and Catalytic Production of Free CO and CN−: Implications for [FeFe]-Hydrogenase Maturation

2021 ◽  
Author(s):  
Eric M. Shepard ◽  
Stella Impano ◽  
Benjamin R. Duffus ◽  
Adrien Pagnier ◽  
Kaitlin S. Duschene ◽  
...  
Keyword(s):  
Hydrogenase Maturation

The organometallic H-cluster of the [FeFe]-hydrogenase consists of a [4Fe-4S] cubane bridged via a cysteinyl thiolate to a 2Fe subcluster ([2Fe]H) containing CO, CN–, and dithiomethylamine (DTMA) ligands. The H-cluster...

scholarly journals Functional Studies of [FeFe] Hydrogenase Maturation in an Escherichia coli Biosynthetic System

2006 ◽  
Vol 188 (6) ◽  
pp. 2163-2172 ◽  
Author(s):  
Paul W. King ◽  
Matthew C. Posewitz ◽  
Maria L. Ghirardi ◽  
Michael Seibert
Keyword(s):  
Escherichia Coli ◽  
Catalytic Domain ◽  
Expression System ◽  
Iron Sulfur Cluster ◽  
Sulfur Cluster ◽  
E Coli ◽  
Functional Studies ◽  
Hydrogenase Maturation ◽  
Iron Sulfur ◽  
And Function

ABSTRACT Maturation of [FeFe] hydrogenases requires the biosynthesis and insertion of the catalytic iron-sulfur cluster, the H cluster. Two radical S-adenosylmethionine (SAM) proteins proposed to function in H cluster biosynthesis, HydEF and HydG, were recently identified in the hydEF-1 mutant of the green alga Chlamydomonas reinhardtii (M. C. Posewitz, P. W. King, S. L. Smolinski, L. Zhang, M. Seibert, and M. L. Ghirardi, J. Biol. Chem. 279:25711-25720, 2004). Previous efforts to study [FeFe] hydrogenase maturation in Escherichia coli by coexpression of C. reinhardtii HydEF and HydG and the HydA1 [FeFe] hydrogenase were hindered by instability of the hydEF and hydG expression clones. A more stable [FeFe] hydrogenase expression system has been achieved in E. coli by cloning and coexpression of hydE, hydF, and hydG from the bacterium Clostridium acetobutylicum. Coexpression of the C. acetobutylicum maturation proteins with various algal and bacterial [FeFe] hydrogenases in E. coli resulted in purified enzymes with specific activities that were similar to those of the enzymes purified from native sources. In the case of structurally complex [FeFe] hydrogenases, maturation of the catalytic sites could occur in the absence of an accessory iron-sulfur cluster domain. Initial investigations of the structure and function of the maturation proteins HydE, HydF, and HydG showed that the highly conserved radical-SAM domains of both HydE and HydG and the GTPase domain of HydF were essential for achieving biosynthesis of active [FeFe] hydrogenases. Together, these results demonstrate that the catalytic domain and a functionally complete set of Hyd maturation proteins are fundamental to achieving biosynthesis of catalytic [FeFe] hydrogenases.

Probing the Solvent Accessibility of the [4Fe–4S] Cluster of the Hydrogenase Maturation Protein HydF from Thermotoga neapolitana by HYSCORE and 3p-ESEEM

2015 ◽  
Vol 119 (43) ◽  
pp. 13680-13689 ◽  
Author(s):  
Marco Albertini ◽  
Paola Berto ◽  
Francesca Vallese ◽  
Marilena Di Valentin ◽  
Paola Costantini ◽  
...  
Keyword(s):  
Solvent Accessibility ◽  
Thermotoga Neapolitana ◽  
Hydrogenase Maturation ◽  
Maturation Protein

scholarly journals The final steps of [FeFe]-hydrogenase maturation

2019 ◽  
Vol 116 (32) ◽  
pp. 15802-15810 ◽  
Author(s):  
Oliver Lampret ◽  
Julian Esselborn ◽  
Rieke Haas ◽  
Andreas Rutz ◽  
Rosalind L. Booth ◽  
...  
Keyword(s):  
Functional Integration ◽  
Structural Features ◽  
Site Directed Mutagenesis ◽  
Structural Reorganization ◽  
Biologically Inspired ◽  
Hydrogenase Maturation ◽  
Flexible Loop ◽  
Loop Region ◽  
Shed Light ◽  
Secondary Ligand

The active site (H-cluster) of [FeFe]-hydrogenases is a blueprint for the design of a biologically inspired H2-producing catalyst. The maturation process describes the preassembly and uptake of the unique [2FeH] cluster into apo-hydrogenase, which is to date not fully understood. In this study, we targeted individual amino acids by site-directed mutagenesis in the [FeFe]-hydrogenase CpI of Clostridium pasteurianum to reveal the final steps of H-cluster maturation occurring within apo-hydrogenase. We identified putative key positions for cofactor uptake and the subsequent structural reorganization that stabilizes the [2FeH] cofactor in its functional coordination sphere. Our results suggest that functional integration of the negatively charged [2FeH] precursor requires the positive charges and individual structural features of the 2 basic residues of arginine 449 and lysine 358, which mark the entrance and terminus of the maturation channel, respectively. The results obtained for 5 glycine-to-histidine exchange variants within a flexible loop region provide compelling evidence that the glycine residues function as hinge positions in the refolding process, which closes the secondary ligand sphere of the [2FeH] cofactor and the maturation channel. The conserved structural motifs investigated here shed light on the interplay between the secondary ligand sphere and catalytic cofactor.

In vitro activation of [FeFe] hydrogenase: new insights into hydrogenase maturation

2007 ◽  
Vol 12 (4) ◽  
pp. 443-447 ◽  
Author(s):  
Shawn E. McGlynn ◽  
Shane S. Ruebush ◽  
Anatoli Naumov ◽  
Lauren E. Nagy ◽  
Alexandra Dubini ◽  
...  
Keyword(s):  
Hydrogenase Maturation ◽  
In Vitro Activation

HypD Is the Scaffold Protein for Fe-(CN)2CO Cofactor Assembly in [NiFe]-Hydrogenase Maturation

Biochemistry ◽  
2013 ◽  
Vol 52 (19) ◽  
pp. 3289-3296 ◽  
Author(s):  
Sven T. Stripp ◽  
Basem Soboh ◽  
Ute Lindenstrauss ◽  
Mario Braussemann ◽  
Martin Herzberg ◽  
...  
Keyword(s):  
Scaffold Protein ◽  
Hydrogenase Maturation

scholarly journals A model system for [NiFe] hydrogenase maturation studies: Purification of an active site-containing hydrogenase large subunit without small subunit

FEBS Letters ◽  
2005 ◽  
Vol 579 (20) ◽  
pp. 4292-4296 ◽  
Author(s):  
Gordon Winter ◽  
Thorsten Buhrke ◽  
Oliver Lenz ◽  
Anne Katherine Jones ◽  
Michael Forgber ◽  
...  
Keyword(s):  
Active Site ◽  
Large Subunit ◽  
Small Subunit ◽  
Model System ◽  
Hydrogenase Maturation

scholarly journals The Maturation Pathway of Nickel Urease

Inorganics ◽  
2019 ◽  
Vol 7 (7) ◽  
pp. 85 ◽  
Author(s):  
Yap Shing Nim ◽  
Kam-Bo Wong
Keyword(s):  
Protein Interactions ◽  
Protein Complexes ◽  
Toxic Metal ◽  
Specific Protein ◽  
Accessory Proteins ◽  
Protein Protein Interactions ◽  
Nickel Ions ◽  
Hydrogenase Maturation ◽  
Maturation Factor

Maturation of urease involves post-translational insertion of nickel ions to form an active site with a carbamylated lysine ligand and is assisted by urease accessory proteins UreD, UreE, UreF and UreG. Here, we review our current understandings on how these urease accessory proteins facilitate the urease maturation. The urease maturation pathway involves the transfer of Ni2+ from UreE → UreG → UreF/UreD → urease. To avoid the release of the toxic metal to the cytoplasm, Ni2+ is transferred from one urease accessory protein to another through specific protein–protein interactions. One central theme depicts the role of guanosine triphosphate (GTP) binding/hydrolysis in regulating the binding/release of nickel ions and the formation of the protein complexes. The urease and [NiFe]-hydrogenase maturation pathways cross-talk with each other as UreE receives Ni2+ from hydrogenase maturation factor HypA. Finally, the druggability of the urease maturation pathway is reviewed.

Sign in / Sign up

Export Citation Format

Share Document