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

Kowalska, Joanna K.; Henthorn, Justin T.; Van Stappen, Casey; Trncik, Christian; Einsle, Oliver; Keavney, David; DeBeer, Serena

doi:10.1002/anie.201901899

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

Abstract

Abstract Nitrogenase enzymes catalyze the reduction of atmospheric dinitrogen to ammonia utilizing a Mo‐7Fe‐9S‐C active site, the so‐called FeMoco cluster. FeMoco and an analogous small‐molecule (Et ₄ N)[(Tp)MoFe ₃ S ₄ Cl ₃ ] cubane have both been proposed to contain unusual spin‐coupled Mo ^III sites with an S (Mo)=1/2 non‐Hund configuration at the Mo atom. Herein, we present Fe and Mo L ₃ ‐edge X‐ray magnetic circular dichroism (XMCD) spectroscopy of the (Et ₄ N)[(Tp)MoFe ₃ S ₄ Cl ₃ ] cubane and Fe L _2,3 ‐edge XMCD spectroscopy of the MoFe protein (containing both FeMoco and the 8Fe‐7S P‐cluster active sites). As the P‐clusters of MoFe protein have an S =0 total spin, these are effectively XMCD‐silent at low temperature and high magnetic field, allowing for FeMoco to be selectively probed by Fe L _2,3 ‐edge XMCD within the intact MoFe protein. Further, Mo L ₃ ‐edge XMCD spectroscopy of the cubane model has provided experimental support for a local S (Mo)=1/2 configuration, demonstrating the power and selectivity of XMCD.

Authors:

^[1]; Henthorn, Justin T. ^[1]; Van Stappen, Casey ^[1]; Trncik, Christian ^[2]; Einsle, Oliver ^[2]; Keavney, David ^[3];

^[1]

Inorganic Spectroscopy Max Planck Institute for Chemical Energy Conversion Stiftstrasse 34–36 45470 Mülheim an der Ruhr Germany
Institute for Biochemistry and BIOSS Centre for Biological Signalling Studies Albert Ludwigs University of Freiburg Albertstrasse 21 79104 Freiburg Germany
Argonne National Laboratory 9700 S. Cass Ave Argonne IL 60439 USA

Publication Date:: Tue Jun 18 00:00:00 EDT 2019

Research Org.:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Org.:: European Commission - Community Research and Development Information Service (CORDIS) - Seventh Framework Programme (FP7); German Research Foundation (DFG); USDOE Office of Science (SC), Basic Energy Sciences (BES)

OSTI Identifier:: 1605471

Alternate Identifier(s):: OSTI ID: 1526781; OSTI ID: 1559977

Grant/Contract Number:: AC02-06CH11357

Resource Type:: Published Article

Journal Name:: Angewandte Chemie (International Edition)

Additional Journal Information:: Journal Name: Angewandte Chemie (International Edition) Journal Volume: 58 Journal Issue: 28; Journal ID: ISSN 1433-7851

Publisher:: Wiley Blackwell (John Wiley & Sons)

Country of Publication:: Germany

Language:: English

Subject:: 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; FeMo cofactor; X-ray magnetic circular dichroism spectroscopy; nitrogen fixation; nitrogenase; spin coupling

Citation Formats


                    Kowalska, Joanna K., Henthorn, Justin T., Van Stappen, Casey, Trncik, Christian, Einsle, Oliver, Keavney, David, and DeBeer, Serena. X‐ray Magnetic Circular Dichroism Spectroscopy Applied to Nitrogenase and Related Models: Experimental Evidence for a Spin‐Coupled Molybdenum(III) Center.  Germany: N. p., 2019. 
Web.  doi:10.1002/anie.201901899.

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                    Kowalska, Joanna K., Henthorn, Justin T., Van Stappen, Casey, Trncik, Christian, Einsle, Oliver, Keavney, David, & DeBeer, Serena. X‐ray Magnetic Circular Dichroism Spectroscopy Applied to Nitrogenase and Related Models: Experimental Evidence for a Spin‐Coupled Molybdenum(III) Center.  Germany.  https://doi.org/10.1002/anie.201901899

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                    Kowalska, Joanna K., Henthorn, Justin T., Van Stappen, Casey, Trncik, Christian, Einsle, Oliver, Keavney, David, and DeBeer, Serena. Tue .  
"X‐ray Magnetic Circular Dichroism Spectroscopy Applied to Nitrogenase and Related Models: Experimental Evidence for a Spin‐Coupled Molybdenum(III) Center".  Germany.  https://doi.org/10.1002/anie.201901899.

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@article{osti_1605471,

  title        = {X‐ray Magnetic Circular Dichroism Spectroscopy Applied to Nitrogenase and Related Models: Experimental Evidence for a Spin‐Coupled Molybdenum(III) Center},

  author       = {Kowalska, Joanna K. and Henthorn, Justin T. and Van Stappen, Casey and Trncik, Christian and Einsle, Oliver and Keavney, David and DeBeer, Serena},

  abstractNote = {Abstract Nitrogenase enzymes catalyze the reduction of atmospheric dinitrogen to ammonia utilizing a Mo‐7Fe‐9S‐C active site, the so‐called FeMoco cluster. FeMoco and an analogous small‐molecule (Et 4 N)[(Tp)MoFe 3 S 4 Cl 3 ] cubane have both been proposed to contain unusual spin‐coupled Mo III sites with an S (Mo)=1/2 non‐Hund configuration at the Mo atom. Herein, we present Fe and Mo L 3 ‐edge X‐ray magnetic circular dichroism (XMCD) spectroscopy of the (Et 4 N)[(Tp)MoFe 3 S 4 Cl 3 ] cubane and Fe L 2,3 ‐edge XMCD spectroscopy of the MoFe protein (containing both FeMoco and the 8Fe‐7S P‐cluster active sites). As the P‐clusters of MoFe protein have an S =0 total spin, these are effectively XMCD‐silent at low temperature and high magnetic field, allowing for FeMoco to be selectively probed by Fe L 2,3 ‐edge XMCD within the intact MoFe protein. Further, Mo L 3 ‐edge XMCD spectroscopy of the cubane model has provided experimental support for a local S (Mo)=1/2 configuration, demonstrating the power and selectivity of XMCD.},

  doi          = {10.1002/anie.201901899},

  journal      = {Angewandte Chemie (International Edition)},

  number       = 28,

  volume       = 58,

  place        = {Germany},

  year         = {Tue Jun 18 00:00:00 EDT 2019},

  month        = {Tue Jun 18 00:00:00 EDT 2019}

}

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

Abstract

Citation Formats

The role of X-ray spectroscopy in understanding the geometric and electronic structure of nitrogenase journal, June 2015

Investigation of CO bound to inhibited forms of nitrogenase MoFe protein by 13C ENDOR journal, August 1995

A Practical Guide to High-resolution X-ray Spectroscopic Measurements and their Applications in Bioinorganic Chemistry journal, July 2016

Nitrogenase. VIII. Mössbauer and EPR spectroscopy. The MoFe protein component from Azotobacter vinelandii OP journal, July 1975

Electron Paramagnetic Resonance of Nitrogenase and Nitrogenase Components from Clostridium pasteurianum W5 and Azotobacter vinelandii OP journal, November 1972

Structural Basis of Biological Nitrogen Fixation journal, January 1996

Structural models for the metal centers in the nitrogenase molybdenum-iron protein journal, September 1992

Competitive cobalt for zinc substitution in mammalian methionine sulfoxide reductase B1 overexpressed in E. coli: structural and functional insight journal, November 2013

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

Iron L 2,3 -Edge X-ray Absorption and X-ray Magnetic Circular Dichroism Studies of Molecular Iron Complexes with Relevance to the FeMoco and FeVco Active Sites of Nitrogenase journal, June 2017

Natural widths of atomic K and L levels, K α X‐ray lines and several K L L Auger lines journal, April 1979

Iron-sulfur clusters in the molybdenum-iron protein component of nitrogenase. Electron paramagnetic resonance of the carbon monoxide inhibited state journal, October 1979

EXAFS studies on the PN and POX states of the P-clusters in nitrogenase journal, August 1998

Reductive Elimination of H 2 Activates Nitrogenase to Reduce the N≡N Triple Bond: Characterization of the E 4 (4H) Janus Intermediate in Wild-Type Enzyme journal, August 2016

Structure−Function Relationships of Alternative Nitrogenases journal, January 1996

New structural insights into the iron-molybdenum cofactor from Azotobacter vinelandii nitrogenase through sulfur K and molybdenum L x-ray absorption edge studies journal, June 1988

Comparative electronic structures of nitrogenase FeMoco and FeVco journal, January 2017

Metal-Ion Valencies of the FeMo Cofactor in CO-Inhibited and Resting State Nitrogenase by 57 Fe Q-Band ENDOR journal, November 1997

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

P-cluster maturation on nitrogenase MoFe protein journal, June 2007

Characterization of Isolated Nitrogenase FeVco journal, September 2010

The molybdenum site of nitrogenase. 2. A comparative study of molybdenum-iron proteins and the iron-molybdenum cofactor by x-ray absorption spectroscopy journal, June 1978

Structural insights into a protein-bound iron-molybdenum cofactor precursor journal, January 2006

Fluorescence detected soft XAS and MCD - applications to bioinorganic systems journal, May 1996

Substrate Interactions with the Nitrogenase Active Site journal, March 2005

Heterometal Cubane-Type MFe 3 S 4 Clusters (M = Mo, V) Trigonally Symmetrized with Hydrotris(pyrazolyl)borate(1−) and Tris(pyrazolyl)methanesulfonate(1−) Capping Ligands journal, February 2002

Crystallographic structure and functional implications of the nitrogenase molybdenum–iron protein from Azotobacter vinelandii journal, December 1992

The iron-molybdenum cofactor of nitrogenase journal, December 1990

Trapping an Intermediate of Dinitrogen (N 2 ) Reduction on Nitrogenase journal, September 2009

Detection of EPR signals assigned to the 1-equiv-oxidized P-clusters of the nitrogenase MoFe-protein from Azotobacter vinelandii journal, October 1993

Testing if the Interstitial Atom, X , of the Nitrogenase Molybdenum−Iron Cofactor Is N or C: ENDOR, ESEEM, and DFT Studies of the S = 3 / 2 Resting State in Multiple Environments journal, December 2007

Identification of a spin-coupled Mo( iii ) in the nitrogenase iron–molybdenum cofactor journal, January 2014

14 N Electron Spin−Echo Envelope Modulation of the S = 3 / 2 Spin System of the Azotobacter vinelandii Nitrogenase Iron−Molybdenum Cofactor † journal, September 1998

Catalysis-dependent selenium incorporation and migration in the nitrogenase active site iron-molybdenum cofactor journal, December 2015

Exploring Electron/Proton Transfer and Conformational Changes in the Nitrogenase MoFe Protein and FeMo-cofactor Through Cryoreduction/EPR Measurements journal, July 2016

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

ENDOR of the resting state of nitrogenase molybdenum-iron proteins from Azotobacter vinelandii, Klebsiella pneumoniae, and Clostridium pasteurianum. Proton, iron-57, molybdenum-95, and sulfur-33 studies journal, June 1986

Core Level Spectroscopy of Solids book, January 2008

STRUCTURE: Nitrogenase Reveals Its Inner Secrets journal, September 2002

The molybdenum site of nitrogenase. Preliminary structural evidence from x-ray absorption spectroscopy journal, May 1978

Mechanism of Molybdenum Nitrogenase journal, January 1996

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

A novel S = 3/2 EPR signal associated with native Fe-proteins of nitrogenase journal, September 1985

X-ray magnetic circular dichroism—a high energy probe of magnetic properties journal, January 2005

Characterization of the metal clusters in the nitrogenase molybdenum-iron and vanadium-iron proteins of Azotobacter vinelandii using magnetic circular dichroism spectroscopy journal, April 1987

X-ray Absorption and Emission Spectroscopic Studies of [L 2 Fe 2 S 2 ] n Model Complexes: Implications for the Experimental Evaluation of Redox States in Iron–Sulfur Clusters journal, April 2016

Nitrogenase and homologs journal, December 2014

Identification of the CO-Binding Cluster in Nitrogenase MoFe Protein by ENDOR of 57 Fe Isotopomers journal, January 1996

High-resolution molybdenum K-edge X-ray absorption spectroscopy analyzed with time-dependent density functional theory journal, January 2013

Molybdenum L-Edge XAS Spectra of MoFe Nitrogenase: Molybdenum L-Edge XAS Spectra of MoFe Nitrogenase journal, November 2014

Decoding the Nitrogenase Mechanism: The Homologue Approach journal, March 2010

Nitrogenase FeMoco investigated by spatially resolved anomalous dispersion refinement journal, March 2016

The role of X-ray spectroscopy in understanding the geometric and electronic structure of nitrogenase
journal, June 2015

Investigation of CO bound to inhibited forms of nitrogenase MoFe protein by 13C ENDOR
journal, August 1995

A Practical Guide to High-resolution X-ray Spectroscopic Measurements and their Applications in Bioinorganic Chemistry
journal, July 2016

Nitrogenase. VIII. Mössbauer and EPR spectroscopy. The MoFe protein component from Azotobacter vinelandii OP
journal, July 1975

Electron Paramagnetic Resonance of Nitrogenase and Nitrogenase Components from Clostridium pasteurianum W5 and Azotobacter vinelandii OP
journal, November 1972

Structural Basis of Biological Nitrogen Fixation
journal, January 1996

Structural models for the metal centers in the nitrogenase molybdenum-iron protein
journal, September 1992

Competitive cobalt for zinc substitution in mammalian methionine sulfoxide reductase B1 overexpressed in E. coli: structural and functional insight
journal, November 2013

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

Iron L _2,3 -Edge X-ray Absorption and X-ray Magnetic Circular Dichroism Studies of Molecular Iron Complexes with Relevance to the FeMoco and FeVco Active Sites of Nitrogenase
journal, June 2017

Natural widths of atomic K and L levels, K α X‐ray lines and several K L L Auger lines
journal, April 1979

Iron-sulfur clusters in the molybdenum-iron protein component of nitrogenase. Electron paramagnetic resonance of the carbon monoxide inhibited state
journal, October 1979

EXAFS studies on the PN and POX states of the P-clusters in nitrogenase
journal, August 1998

Reductive Elimination of H ₂ Activates Nitrogenase to Reduce the N≡N Triple Bond: Characterization of the E ₄ (4H) Janus Intermediate in Wild-Type Enzyme
journal, August 2016

Structure−Function Relationships of Alternative Nitrogenases
journal, January 1996

New structural insights into the iron-molybdenum cofactor from Azotobacter vinelandii nitrogenase through sulfur K and molybdenum L x-ray absorption edge studies
journal, June 1988

Comparative electronic structures of nitrogenase FeMoco and FeVco
journal, January 2017

Metal-Ion Valencies of the FeMo Cofactor in CO-Inhibited and Resting State Nitrogenase by ⁵⁷ Fe Q-Band ENDOR
journal, November 1997

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

P-cluster maturation on nitrogenase MoFe protein
journal, June 2007

Characterization of Isolated Nitrogenase FeVco
journal, September 2010

The molybdenum site of nitrogenase. 2. A comparative study of molybdenum-iron proteins and the iron-molybdenum cofactor by x-ray absorption spectroscopy
journal, June 1978

Structural insights into a protein-bound iron-molybdenum cofactor precursor
journal, January 2006

Fluorescence detected soft XAS and MCD - applications to bioinorganic systems
journal, May 1996

Substrate Interactions with the Nitrogenase Active Site
journal, March 2005

Heterometal Cubane-Type MFe ₃ S ₄ Clusters (M = Mo, V) Trigonally Symmetrized with Hydrotris(pyrazolyl)borate(1−) and Tris(pyrazolyl)methanesulfonate(1−) Capping Ligands
journal, February 2002

Crystallographic structure and functional implications of the nitrogenase molybdenum–iron protein from Azotobacter vinelandii
journal, December 1992

The iron-molybdenum cofactor of nitrogenase
journal, December 1990

Trapping an Intermediate of Dinitrogen (N ₂ ) Reduction on Nitrogenase
journal, September 2009

Detection of EPR signals assigned to the 1-equiv-oxidized P-clusters of the nitrogenase MoFe-protein from Azotobacter vinelandii
journal, October 1993

Testing if the Interstitial Atom, X , of the Nitrogenase Molybdenum−Iron Cofactor Is N or C: ENDOR, ESEEM, and DFT Studies of the S = ³ / ₂ Resting State in Multiple Environments
journal, December 2007

Identification of a spin-coupled Mo( iii ) in the nitrogenase iron–molybdenum cofactor
journal, January 2014

¹⁴ N Electron Spin−Echo Envelope Modulation of the S = ³ / ₂ Spin System of the Azotobacter vinelandii Nitrogenase Iron−Molybdenum Cofactor ^†
journal, September 1998

Catalysis-dependent selenium incorporation and migration in the nitrogenase active site iron-molybdenum cofactor
journal, December 2015

Exploring Electron/Proton Transfer and Conformational Changes in the Nitrogenase MoFe Protein and FeMo-cofactor Through Cryoreduction/EPR Measurements
journal, July 2016

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

ENDOR of the resting state of nitrogenase molybdenum-iron proteins from Azotobacter vinelandii, Klebsiella pneumoniae, and Clostridium pasteurianum. Proton, iron-57, molybdenum-95, and sulfur-33 studies
journal, June 1986

Core Level Spectroscopy of Solids
book, January 2008

STRUCTURE: Nitrogenase Reveals Its Inner Secrets
journal, September 2002

The molybdenum site of nitrogenase. Preliminary structural evidence from x-ray absorption spectroscopy
journal, May 1978

Mechanism of Molybdenum Nitrogenase
journal, January 1996

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

A novel S = 3/2 EPR signal associated with native Fe-proteins of nitrogenase
journal, September 1985

X-ray magnetic circular dichroism—a high energy probe of magnetic properties
journal, January 2005

Characterization of the metal clusters in the nitrogenase molybdenum-iron and vanadium-iron proteins of Azotobacter vinelandii using magnetic circular dichroism spectroscopy
journal, April 1987

X-ray Absorption and Emission Spectroscopic Studies of [L ₂ Fe ₂ S ₂ ] ⁿ Model Complexes: Implications for the Experimental Evaluation of Redox States in Iron–Sulfur Clusters
journal, April 2016

Nitrogenase and homologs
journal, December 2014

Identification of the CO-Binding Cluster in Nitrogenase MoFe Protein by ENDOR of ⁵⁷ Fe Isotopomers
journal, January 1996

High-resolution molybdenum K-edge X-ray absorption spectroscopy analyzed with time-dependent density functional theory
journal, January 2013

Molybdenum L-Edge XAS Spectra of MoFe Nitrogenase: Molybdenum L-Edge XAS Spectra of MoFe Nitrogenase
journal, November 2014

Decoding the Nitrogenase Mechanism: The Homologue Approach
journal, March 2010

Nitrogenase FeMoco investigated by spatially resolved anomalous dispersion refinement
journal, March 2016