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Title: Electron Spin Relaxation and Biochemical Characterization of the Hydrogenase Maturase HydF: Insights into [2Fe-2S] and [4Fe-4S] Cluster Communication and Hydrogenase Activation

Authors:
; ; ; ; ; ; ; ; ; ORCiD logo
Publication Date:
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
OSTI Identifier:
1363692
Grant/Contract Number:
FG02-10ER16194
Resource Type:
Journal Article: Published Article
Journal Name:
Biochemistry
Additional Journal Information:
Related Information: CHORUS Timestamp: 2017-11-27 06:55:56; Journal ID: ISSN 0006-2960
Publisher:
American Chemical Society (ACS)
Country of Publication:
United States
Language:
English

Citation Formats

Shepard, Eric M., Byer, Amanda S., Aggarwal, Priyanka, Betz, Jeremiah N., Scott, Anna G., Shisler, Krista A., Usselman, Robert J., Eaton, Gareth R., Eaton, Sandra S., and Broderick, Joan B.. Electron Spin Relaxation and Biochemical Characterization of the Hydrogenase Maturase HydF: Insights into [2Fe-2S] and [4Fe-4S] Cluster Communication and Hydrogenase Activation. United States: N. p., 2017. Web. doi:10.1021/acs.biochem.7b00169.
Shepard, Eric M., Byer, Amanda S., Aggarwal, Priyanka, Betz, Jeremiah N., Scott, Anna G., Shisler, Krista A., Usselman, Robert J., Eaton, Gareth R., Eaton, Sandra S., & Broderick, Joan B.. Electron Spin Relaxation and Biochemical Characterization of the Hydrogenase Maturase HydF: Insights into [2Fe-2S] and [4Fe-4S] Cluster Communication and Hydrogenase Activation. United States. doi:10.1021/acs.biochem.7b00169.
Shepard, Eric M., Byer, Amanda S., Aggarwal, Priyanka, Betz, Jeremiah N., Scott, Anna G., Shisler, Krista A., Usselman, Robert J., Eaton, Gareth R., Eaton, Sandra S., and Broderick, Joan B.. 2017. "Electron Spin Relaxation and Biochemical Characterization of the Hydrogenase Maturase HydF: Insights into [2Fe-2S] and [4Fe-4S] Cluster Communication and Hydrogenase Activation". United States. doi:10.1021/acs.biochem.7b00169.
@article{osti_1363692,
title = {Electron Spin Relaxation and Biochemical Characterization of the Hydrogenase Maturase HydF: Insights into [2Fe-2S] and [4Fe-4S] Cluster Communication and Hydrogenase Activation},
author = {Shepard, Eric M. and Byer, Amanda S. and Aggarwal, Priyanka and Betz, Jeremiah N. and Scott, Anna G. and Shisler, Krista A. and Usselman, Robert J. and Eaton, Gareth R. and Eaton, Sandra S. and Broderick, Joan B.},
abstractNote = {},
doi = {10.1021/acs.biochem.7b00169},
journal = {Biochemistry},
number = ,
volume = ,
place = {United States},
year = 2017,
month = 5
}

Journal Article:
Free Publicly Available Full Text
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  • The in vitro activation of the [FeFe] hydrogenase is accomplished by combining Escherichia coli cell extracts containing the heterologously expressed inactive HydA with extracts in which hydrogenase-specific maturation proteins HydE, HydF, and HydG are expressed in concert. Interestingly, the process of HydA activation occurs rapidly and in the absence of potential substrates, which suggests that the hydrogenase accessory proteins synthesize an H-cluster precursor that can be quickly transferred to the hydrogenase enzyme to affect activation. HydA activity is observed to be dependent on the protein fraction containing all three accessory proteins expressed in concert and cannot be accomplished with additionmore » of heat-treated extract or extract filtrate, suggesting that the activation of the hydrogenase structural protein is mediated by interaction with the accessory assembly protein(s). These results represent the first important step in understanding the process of H-cluster assembly and provide significant insights into hydrogenase maturation.« less
  • A conserved cysteine located in the signature motif of the catalytic center (H-cluster) of [FeFe]-hydrogenases functions in proton transfer. This residue corresponds to C298 in Clostridium acetobutylicum CaHydA. Despite the chemical and structural difference, the mutant C298D retains fast catalytic activity, while replacement with any other amino acid caused significant activity loss. Given the proximity of C298 to the H-cluster, the effect of the C298D mutation on the catalytic center was studied by continuous wave (CW) and pulse electron paramagnetic resonance (EPR) and by Fourier transform infrared (FTIR) spectroscopies. Comparison of the C298D mutant with the wild type CaHydA bymore » CW and pulse EPR showed that the electronic structure of the center is not altered. FTIR spectroscopy confirmed that absorption peak values observed in the mutant are virtually identical to those observed in the wild type, indicating that the H-cluster is not generally affected by the mutation. Significant differences were observed only in the inhibited state Hox-CO: the vibrational modes assigned to the COexo and Fed-CO in this state are shifted to lower values in C298D, suggesting different interaction of these ligands with the protein moiety when C298 is changed to D298. More relevant to the catalytic cycle, the redox equilibrium between the Hox and Hred states is modified by the mutation, causing a prevalence of the oxidized state. This work highlights how the interactions between the protein environment and the H-cluster, a dynamic closely interconnected system, can be engineered and studied in the perspective of designing bio-inspired catalysts and mimics.« less
  • We report analytical calculations of isotropic hyperfine-coupling constants in radicals using a spin-adapted open-shell coupled-cluster theory, namely, the unitary group based combinatoric open-shell coupled-cluster (COSCC) approach within the singles and doubles approximation. A scheme for the evaluation of the one-particle spin-density matrix required in these calculations is outlined within the spin-free formulation of the COSCC approach. In this scheme, the one-particle spin-density matrix for an open-shell state with spin S and M{sub S} = + S is expressed in terms of the one- and two-particle spin-free (charge) density matrices obtained from the Lagrangian formulation that is used for calculating themore » analytic first derivatives of the energy. Benchmark calculations are presented for NO, NCO, CH{sub 2}CN, and two conjugated π-radicals, viz., allyl and 1-pyrrolyl in order to demonstrate the performance of the proposed scheme.« less