Search Menu
DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information
Search Scholarly Publications

Title: Exploring Electron/Proton Transfer and Conformational Changes in the Nitrogenase MoFe Protein and FeMo-cofactor Through Cryoreduction/EPR Measurements

Abstract

Here, we combine cryoreduction/annealing/EPR measurements of nitrogenase MoFe protein with results of earlier investigations to provide a detailed view of the electron/proton transfer events and conformational changes that occur during early stages of [e-/H+] accumulation by the MoFe protein. This includes reduction of (i) the non-catalytic state of the iron-molybdenum cofactor (FeMo-co) active site that is generated by chemical oxidation of the resting-state cofactor (S = 3/2)) within resting MoFe (E0), and (ii) the catalytic state that has accumulated n =1 [e-/H+] above the resting-state level, denoted E1(1H) (S ≥ 1) in the Lowe-Thorneley kinetic scheme. FeMo-co does not undergo a major change of conformation during reduction of oxidized FeMo-co. In contrast, FeMo-co undergoes substantial conformational changes during the reduction of E0 to E1(1H), and of E1(1H) to E2(2H) (n = 2; S = 3/2). The experimental results further suggest that the E1(1H) → E2(2H) step involves coupled delivery of a proton and electron (PCET) to FeMo-co of E1(H) to generate a non-equilibrium S = ½ form E2(2H)*. This subsequently undergoes conformational relaxation and attendant change in FeMo-co spin state, to generate the equilibrium E2(2H) (S = 3/2) state. Unexpectedly, these experiments also reveal conformational coupling between FeMo-co and P-cluster,more » and between Fe protein binding and FeMo-co, which might play a role in gated ET from reduced Fe protein to FeMo-co.« less

Authors:
 [1];  [2];  [2];  [2];  [1];  [3];  [2];  [1]
+ Show Author Affiliations
  1. Northwestern Univ., Evanston, IL (United States). Dept. of Chemistry
  2. Utah State Univ., Logan, UT (United States). Dept. of Chemistry and Biochemistry
  3. Virginia Polytechnic Inst. and State Univ. (Virginia Tech), Blacksburg, VA (United States). Dept. of Biochemistry
Publication Date:
Research Org.:
Utah State Univ., Logan, UT (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES); National Institutes of Health (NIH)
OSTI Identifier:
1463083
Alternate Identifier(s):
OSTI ID: 1401224
Grant/Contract Number:  
SC0010687; GM 111097; SC0010834
Resource Type:
Accepted Manuscript
Journal Name:
Israel Journal of Chemistry
Additional Journal Information:
Journal Volume: 56; Journal Issue: 9-10; Journal ID: ISSN 0021-2148
Publisher:
Wiley
Country of Publication:
United States
Language:
English
Subject:
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; 59 BASIC BIOLOGICAL SCIENCES; conformation analysis; electron transfer; enzyme catalysis; EPR spectroscopy; proton transfer

Citation Formats

Davydov, Roman, Khadka, Nimesh, Yang, Zhi-Yong, Fielding, Andrew J., Lukoyanov, Dmitriy, Dean, Dennis R., Seefeldt, Lance C., and Hoffman, Brian M. Exploring Electron/Proton Transfer and Conformational Changes in the Nitrogenase MoFe Protein and FeMo-cofactor Through Cryoreduction/EPR Measurements. United States: N. p., 2016. Web. doi:10.1002/ijch.201600026.
Copy to clipboard
Davydov, Roman, Khadka, Nimesh, Yang, Zhi-Yong, Fielding, Andrew J., Lukoyanov, Dmitriy, Dean, Dennis R., Seefeldt, Lance C., & Hoffman, Brian M. Exploring Electron/Proton Transfer and Conformational Changes in the Nitrogenase MoFe Protein and FeMo-cofactor Through Cryoreduction/EPR Measurements. United States. https://doi.org/10.1002/ijch.201600026
Copy to clipboard
Davydov, Roman, Khadka, Nimesh, Yang, Zhi-Yong, Fielding, Andrew J., Lukoyanov, Dmitriy, Dean, Dennis R., Seefeldt, Lance C., and Hoffman, Brian M. Fri . "Exploring Electron/Proton Transfer and Conformational Changes in the Nitrogenase MoFe Protein and FeMo-cofactor Through Cryoreduction/EPR Measurements". United States. https://doi.org/10.1002/ijch.201600026. https://www.osti.gov/servlets/purl/1463083.
Copy to clipboard
@article{osti_1463083,
title = {Exploring Electron/Proton Transfer and Conformational Changes in the Nitrogenase MoFe Protein and FeMo-cofactor Through Cryoreduction/EPR Measurements},
author = {Davydov, Roman and Khadka, Nimesh and Yang, Zhi-Yong and Fielding, Andrew J. and Lukoyanov, Dmitriy and Dean, Dennis R. and Seefeldt, Lance C. and Hoffman, Brian M.},
abstractNote = {Here, we combine cryoreduction/annealing/EPR measurements of nitrogenase MoFe protein with results of earlier investigations to provide a detailed view of the electron/proton transfer events and conformational changes that occur during early stages of [e-/H+] accumulation by the MoFe protein. This includes reduction of (i) the non-catalytic state of the iron-molybdenum cofactor (FeMo-co) active site that is generated by chemical oxidation of the resting-state cofactor (S = 3/2)) within resting MoFe (E0), and (ii) the catalytic state that has accumulated n =1 [e-/H+] above the resting-state level, denoted E1(1H) (S ≥ 1) in the Lowe-Thorneley kinetic scheme. FeMo-co does not undergo a major change of conformation during reduction of oxidized FeMo-co. In contrast, FeMo-co undergoes substantial conformational changes during the reduction of E0 to E1(1H), and of E1(1H) to E2(2H) (n = 2; S = 3/2). The experimental results further suggest that the E1(1H) → E2(2H) step involves coupled delivery of a proton and electron (PCET) to FeMo-co of E1(H) to generate a non-equilibrium S = ½ form E2(2H)*. This subsequently undergoes conformational relaxation and attendant change in FeMo-co spin state, to generate the equilibrium E2(2H) (S = 3/2) state. Unexpectedly, these experiments also reveal conformational coupling between FeMo-co and P-cluster, and between Fe protein binding and FeMo-co, which might play a role in gated ET from reduced Fe protein to FeMo-co.},
doi = {10.1002/ijch.201600026},
journal = {Israel Journal of Chemistry},
number = 9-10,
volume = 56,
place = {United States},
year = {Fri Jul 29 00:00:00 EDT 2016},
month = {Fri Jul 29 00:00:00 EDT 2016}
}
Copy to clipboard
Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
https://doi.org/10.1002/ijch.201600026
Copyright Statement
Other availability
Search WorldCat Search WorldCat to find libraries that may hold this journal
Citation Metrics:
Cited by: 11 works
Citation information provided by
Web of Science
Save / Share:
Export Metadata
Save to My Library
You must Sign In or Create an Account in order to save documents to your library.

Works referenced in this record:

Unification of reaction pathway and kinetic scheme for N2 reduction catalyzed by nitrogenase
journal, March 2012

  • Lukoyanov, D.; Yang, Z. -Y.; Barney, B. M.
  • Proceedings of the National Academy of Sciences, Vol. 109, Issue 15
  • DOI: 10.1073/pnas.1202197109

Proton Transfer at Helium Temperatures during Dioxygen Activation by Heme Monooxygenases
journal, December 2004

  • Davydov, Roman; Chemerisov, Sergey; Werst, David E.
  • Journal of the American Chemical Society, Vol. 126, Issue 49
  • DOI: 10.1021/ja044646t

Is Mo Involved in Hydride Binding by the Four-Electron Reduced (E 4 ) Intermediate of the Nitrogenase MoFe Protein?
journal, March 2010

  • Lukoyanov, Dmitriy; Yang, Zhi-Yong; Dean, Dennis R.
  • Journal of the American Chemical Society, Vol. 132, Issue 8
  • DOI: 10.1021/ja910613m

USE OF GLASS ELECTRODES TO MEASURE ACIDITIES IN DEUTERIUM OXIDE 1,2
journal, January 1960

  • Glasoe, Paul K.; Long, F. A.
  • The Journal of Physical Chemistry, Vol. 64, Issue 1
  • DOI: 10.1021/j100830a521

Catalytic and Biophysical Properties of a Nitrogenase Apo-MoFe Protein Produced by a n ifB -Deletion Mutant of Azotobacter v inelandii †
journal, September 1998

  • Christiansen, Jason; Goodwin, Paul J.; Lanzilotta, William N.
  • Biochemistry, Vol. 37, Issue 36
  • DOI: 10.1021/bi981165b

Temperature Invariance of the Nitrogenase Electron Transfer Mechanism
journal, October 2012

  • Mayweather, Diana; Danyal, Karamatullah; Dean, Dennis R.
  • Biochemistry, Vol. 51, Issue 42
  • DOI: 10.1021/bi301164j

Electron transfer in nitrogenase catalysis
journal, April 2012

  • Seefeldt, Lance C.; Hoffman, Brian M.; Dean, Dennis R.
  • Current Opinion in Chemical Biology, Vol. 16, Issue 1-2
  • DOI: 10.1016/j.cbpa.2012.02.012

P + State of Nitrogenase P-Cluster Exhibits Electronic Structure of a [Fe 4 S 4 ] + Cluster
journal, August 2012

  • Rupnik, Kresimir; Hu, Yilin; Lee, Chi Chung
  • Journal of the American Chemical Society, Vol. 134, Issue 33
  • DOI: 10.1021/ja304077h

Catalytic conversion of nitrogen to ammonia by an iron model complex
journal, September 2013

  • Anderson, John S.; Rittle, Jonathan; Peters, Jonas C.
  • Nature, Vol. 501, Issue 7465
  • DOI: 10.1038/nature12435

Controlled protonation of iron–molybdenum cofactor by nitrogenase: a structural and theoretical analysis
journal, April 2001

Connecting nitrogenase intermediates with the kinetic scheme for N2 reduction by a relaxation protocol and identification of the N2 binding state
journal, January 2007

  • Lukoyanov, D.; Barney, B. M.; Dean, D. R.
  • Proceedings of the National Academy of Sciences, Vol. 104, Issue 5
  • DOI: 10.1073/pnas.0610975104

Electron-paramagnetic-resonance studies on the redox properties of the molybdenum-iron protein of nitrogenase between +50 and −450 mV
journal, September 1978

  • O'Donnell, M. J.; Smith, B. E.
  • Biochemical Journal, Vol. 173, Issue 3
  • DOI: 10.1042/bj1730831

Mechanism of Nitrogen Fixation by Nitrogenase: The Next Stage
journal, January 2014

  • Hoffman, Brian M.; Lukoyanov, Dmitriy; Yang, Zhi-Yong
  • Chemical Reviews, Vol. 114, Issue 8
  • DOI: 10.1021/cr400641x

Proton-Coupled Electron Transfer
journal, April 2012

  • Weinberg, David R.; Gagliardi, Christopher J.; Hull, Jonathan F.
  • Chemical Reviews, Vol. 112, Issue 7
  • DOI: 10.1021/cr200177j

Nitrogenase and nitrogenase reductase associate and dissociate with each catalytic cycle.
journal, June 1978

  • Hageman, R. V.; Burris, R. H.
  • Proceedings of the National Academy of Sciences, Vol. 75, Issue 6
  • DOI: 10.1073/pnas.75.6.2699

Conformational Gating of Electron Transfer from the Nitrogenase Fe Protein to MoFe Protein
journal, May 2010

  • Danyal, Karamatullah; Mayweather, Diana; Dean, Dennis R.
  • Journal of the American Chemical Society, Vol. 132, Issue 20
  • DOI: 10.1021/ja101737f

Fe and Mo EXAFS of Azotobacter vinelandii Nitrogenase in Partially Oxidized and Singly Reduced Forms
journal, October 1995

  • Christiansen, J.; Tittsworth, R. C.; Hales, B. J.
  • Journal of the American Chemical Society, Vol. 117, Issue 40
  • DOI: 10.1021/ja00145a012

Nitrogenase: A Draft Mechanism
journal, December 2012

  • Hoffman, Brian M.; Lukoyanov, Dmitriy; Dean, Dennis R.
  • Accounts of Chemical Research, Vol. 46, Issue 2
  • DOI: 10.1021/ar300267m

Electron transfer precedes ATP hydrolysis during nitrogenase catalysis
journal, September 2013

  • Duval, S.; Danyal, K.; Shaw, S.
  • Proceedings of the National Academy of Sciences, Vol. 110, Issue 41
  • DOI: 10.1073/pnas.1311218110

Biochemistry and Theory of Proton-Coupled Electron Transfer
journal, January 2014

  • Migliore, Agostino; Polizzi, Nicholas F.; Therien, Michael J.
  • Chemical Reviews, Vol. 114, Issue 7
  • DOI: 10.1021/cr4006654

EPR and ENDOR studies of Fe(II) hemoproteins reduced and oxidized at 77 K
journal, December 2007

  • Davydov, Roman; Hoffman, Brian M.
  • JBIC Journal of Biological Inorganic Chemistry, Vol. 13, Issue 3
  • DOI: 10.1007/s00775-007-0328-5

On reversible H2 loss upon N2 binding to FeMo-cofactor of nitrogenase
journal, September 2013

  • Yang, Z. -Y.; Khadka, N.; Lukoyanov, D.
  • Proceedings of the National Academy of Sciences, Vol. 110, Issue 41
  • DOI: 10.1073/pnas.1315852110

Electron Paramagnetic Resonance and Electron-Nuclear Double Resonance Studies of the Reactions of Cryogenerated Hydroperoxoferric–Hemoprotein Intermediates
journal, July 2014

  • Davydov, Roman; Laryukhin, Mikhail; Ledbetter-Rogers, Amy
  • Biochemistry, Vol. 53, Issue 30
  • DOI: 10.1021/bi500296d

A Confirmation of the Quench-Cryoannealing Relaxation Protocol for Identifying Reduction States of Freeze-Trapped Nitrogenase Intermediates
journal, March 2014

  • Lukoyanov, Dmitriy; Yang, Zhi-Yong; Duval, Simon
  • Inorganic Chemistry, Vol. 53, Issue 7
  • DOI: 10.1021/ic500013c

Evidence for Interstitial Carbon in Nitrogenase FeMo Cofactor
journal, November 2011

Molybdenum-95 and proton ENDOR spectroscopy of the nitrogenase molybdenum-iron protein
journal, February 1982

  • Hoffman, Brian M.; Roberts, James E.; Orme-Johnson, W. H.
  • Journal of the American Chemical Society, Vol. 104, Issue 3
  • DOI: 10.1021/ja00367a042

Q-Band Pulsed Electron Spin-Echo Spectrometer and Its Application to ENDOR and ESEEM
journal, March 1996

  • Davoust, Clark E.; Doan, Peter E.; Hoffman, Brian M.
  • Journal of Magnetic Resonance, Series A, Vol. 119, Issue 1
  • DOI: 10.1006/jmra.1996.0049

Mechanism of Molybdenum Nitrogenase
journal, January 1996

  • Burgess, Barbara K.; Lowe, David J.
  • Chemical Reviews, Vol. 96, Issue 7
  • DOI: 10.1021/cr950055x

X-ray Emission Spectroscopy Evidences a Central Carbon in the Nitrogenase Iron-Molybdenum Cofactor
journal, November 2011

Electron Transfer within Nitrogenase: Evidence for a Deficit-Spending Mechanism
journal, November 2011

  • Danyal, Karamatullah; Dean, Dennis R.; Hoffman, Brian M.
  • Biochemistry, Vol. 50, Issue 43
  • DOI: 10.1021/bi201003a

Active intermediates in heme monooxygenase reactions as revealed by cryoreduction/annealing, EPR/ENDOR studies
journal, March 2011

  • Davydov, Roman; Hoffman, Brian M.
  • Archives of Biochemistry and Biophysics, Vol. 507, Issue 1
  • DOI: 10.1016/j.abb.2010.09.013

Structure of the Nucleotide Radical Formed during Reaction of CDP/TTP with the E441Q-α2β2 of E. coli Ribonucleotide Reductase
journal, January 2009

  • Zipse, Hendrik; Artin, Erin; Wnuk, Stanislaw
  • Journal of the American Chemical Society, Vol. 131, Issue 1
  • DOI: 10.1021/ja806693s

Mössbauer Study of the MoFe Protein of Nitrogenase from Azotobacter vinelandii Using Selective 57 Fe Enrichment of the M-Centers
journal, May 2000

  • Yoo, Sun Jae; Angove, Hayley C.; Papaefthymiou, Vasilios
  • Journal of the American Chemical Society, Vol. 122, Issue 20
  • DOI: 10.1021/ja000254k

Identification of a Key Catalytic Intermediate Demonstrates That Nitrogenase Is Activated by the Reversible Exchange of N 2 for H 2
journal, March 2015

  • Lukoyanov, Dmitriy; Yang, Zhi-Yong; Khadka, Nimesh
  • Journal of the American Chemical Society, Vol. 137, Issue 10
  • DOI: 10.1021/jacs.5b00103

A nitrogen pressure of 50 atmospheres does not prevent evolution of hydrogen by nitrogenase
journal, June 1984

    Sort by:
    [ × clear filter / sort ]

    Works referencing / citing this record:

    X-ray Magnetic Circular Dichroism Spectroscopy Applied to Nitrogenase and Related Models: Experimental Evidence for a Spin-Coupled Molybdenum(III) Center
    journal, June 2019

    • Kowalska, Joanna K.; Henthorn, Justin T.; Van Stappen, Casey
    • Angewandte Chemie International Edition, Vol. 58, Issue 28
    • DOI: 10.1002/anie.201901899

    X‐ray Magnetic Circular Dichroism Spectroscopy Applied to Nitrogenase and Related Models: Experimental Evidence for a Spin‐Coupled Molybdenum(III) Center
    journal, June 2019

    • Kowalska, Joanna K.; Henthorn, Justin T.; Van Stappen, Casey
    • Angewandte Chemie, Vol. 131, Issue 28
    • DOI: 10.1002/ange.201901899
      Sort by:
      [ × clear filter / sort ]

      Similar Records in DOE PAGES and OSTI.GOV collections: