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Published on 30 September 2016 on http://pubs.rsc.org | doi:10.1039/9781782628842-FP001
Molybdenum and Tungsten Enzymes

Spectroscopic and Theoretical Investigations
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5: Molybdenum and Tungsten Enzymes: Biochemistry
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Molybdenum and Tungsten

Enzymes
Spectroscopic and Theoretical Investigations
Edited by
Russ Hille
University of California, Riverside, CA, USA
Email: russ.hille@ucr.edu
Carola Schulzke
University of Greifswald, Germany
Email: carola.schulzke@uni-greifswald.de
and
Martin L. Kirk
University of New Mexico, Albuquerque, NM, USA
Email: mkirk@unm.edu
Published on 30 September 2016 on http://pubs.rsc.org | doi:10.1039/9781782628842-FP001 View Online
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Preface
In the late 1950s and early 1960s, evidence was accumulating that molybde
num was not simply present in the enzyme xanthine oxidase from cow's milk
but that it was required for its activity and changed its oxidation state in the
course of the reaction with substrate. In a tour-de-force isotopic substitution
study reported in Nature in 1966, R.C. Bray and L.S. Meriwether demons
trated unequivocally that the EPR signals elicited by the enzyme upon treat
ment with xanthine arose from a molybdenum-containing active site. It is
a happy coincidence but altogether fitting that this volume marks the 50th
anniversary of this seminal work.
For many years, only five enzymes were recognized as possessing molyb
denum in their active sites: nitrogenase from bacteria such as Klebsiella
pneumoniae and Azotobacter vinelandii; xanthine oxidase from bovine milk
(and other vertebrate sources); aldehyde oxidase from vertebrate as well as
bacterial sources; the vertebrate sulfite oxidase; and the assimilatory nitrate
reductase from plants (and algae and fungi). That began to change in the
1980s with the demonstration by K. V. Rajagopalan that an organic cofac
tor accompanied the molybdenum in the active sites of these enzymes (with
the exception of nitrogenase), and with the contemporaneous discovery that
tungsten was also found in the active sites of enzymes in certain bacteria.
There are now several dozen molybdenum- and tungsten-containing
enzymes that have been crystallographically characterized, along with most
of the enzymes responsible for the biosynthesis of the organic cofactor vari
ously known as molybdopterin, tungstopterin and pyranopterin. The active
site metal centres of these enzymes have proven to be fascinating and chal
lenging targets for synthetic inorganic chemists, and both enzymes and
synthetic models have proven fertile ground for the application of a range
of physicochemical and spectroscopic methods probing their physical and
electronic structures as well as their intrinsic reactivity. At present, well

Molybdenum and Tungsten Enzymes: Spectroscopic and Theoretical Investigations
Edited by Russ Hille, Carola Schulzke, and Martin L. Kirk
Published by the Royal Society of Chemistry, www.rsc.org
v
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vi Preface
over 50 molybdenum- and tungsten-containing enzymes have been isolated
and characterized, and these have been found to catalyze a broad range of
oxidation-reduction reactions, and even reactions that (at least formally) do
not involve oxidation–reduction of substrate. These enzymes are found in
a wide range of metabolic pathways and play particularly prominent roles

in the global cycling of nitrogen, sulfur and carbon. Many have vital roles
in bacterial bioenergetics, catalyzing crucial energy-conserving reactions
under a variety of growth conditions. Indeed, they seem to have been among
the earliest enzyme systems to have arisen, as reflected in their near-universal
distribution in the biosphere. Finally, genomics analyses have led to the
identification of hundreds of genes encoding putative new proteins that are
likely to possess one or another metal. These systems represent an enormous
frontier of new enzymes that remains to be explored.
This title provides an up-to-date account of the state of our understanding
of molybdenum and tungsten enzymes and is divided into three volumes,
dealing with: (1) the enzymes themselves, along with pyranopterin cofac
tor biosynthesis and incorporation of the mature cofactor into apoprotein
(Molybdenum and Tungsten Enzymes: Biochemistry), (2) inorganic complexes
that model the structures and/or reactivity of the active sites of each major
group of molybdenum and tungsten enzymes (Molybdenum and Tungsten
Enzymes: Inorganic Chemistry) and (3) spectroscopic and related methods of
physical chemistry (including computational work) that have been applied
to both enzymes and model compounds (Molybdenum and Tungsten Enzymes:
Physical Methods). Each volume is introduced by an overview chapter written
by a leading expert in the field, followed by the individual chapters that detail
specific topics associated with each volume. The intent of these overview
chapters is to provide an overarching and unifying theme that places each of
the three major subject areas in proper context.
We are deeply indebted to each of the contributors for their efforts, which
lay out the current state of our understanding in each of the many subject
areas considered. The coverage of these volumes is inevitably incomplete
due to space constraints, however, and for this we apologize. However, the
topics that are covered are presented to the reader in considerable detail;
written in a style and spirit that will be fully accessible by current researchers
in the field as well as those who wish to learn more about these fascinating
metalloproteins. We sincerely hope that these volumes will underscore how
rapid the progress has been over the past decade or so, and also how rapidly
the field is expanding. The ultimate goal is to stimulate further research on
molybdenum and tungsten enzymes, and especially to encourage new inves
tigators to take up one or another aspect of these systems. It seems inevitable
that many exciting new discoveries lie in wait.
Russ Hille
Carola Schulzke
Martin L. Kirk
Dedication
It is all too fitting that these volumes dealing with the bioinorganic chemistry
of molybdenum and tungsten be dedicated to three outstanding chemists
whose contributions to the field over many years continues to inform, illumi-
nate and inspire: Richard H. Holm, C. David Garner and John H. Enemark.
Prof. Holm has over 500 research publications (cited over 35 000 times)
covering a wide range of nickel, iron and molybdenum chemistry (among
other transition metals). He is perhaps most widely recognized for studies,
beginning in the 1970s, that describe the synthesis and characterization
of iron-sulfur clusters. This work came to include modelling the M and P
clusters of nitrogenase, which perhaps provided the motivation to investi-
gate models of mononuclear molybdenum-containing enzymes. His molyb-
denum work achieved great success with the synthesis of MoO2 models for
enzymes of the sulfite oxidase, and later the DMSO reductase family, and the
characterization of their properties as oxygen atom transfer catalysts. A key
contribution was his use of bulky ligands to the metal that prevented µ-oxo

vii
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viii Dedication
dimerization, which had long stymied work in the field. He is Higgins Profes-
sor of Chemistry at Harvard University, a member of the National Academy
of Sciences and the recipient of many other awards.
Prof. Garner already had a strong track record in the synthesis of copper
and molybdenum complexes when, beginning in the late 1970s, he became

one of the first researchers to apply the then-new analytical method of X-ray
absorption spectroscopy not only to models of molybdenum enzymes but
also to the enzymes themselves. The discovery of thiolate-like sulfur, Mo=O
and Mo=S ligands to the metal in the active sites of enzymes such as sulfite
oxidase, xanthine oxidase and DMSO reductase was critical in establishing
the molybdenum coordination environment in these enzymes and greatly
focused efforts to synthesize accurate structural and functional mimics of
the enzymes. With over 300 publications (having over 8000 citations), he is
presently Professor Emeritus at the University of Nottingham and a Fellow of
the Royal Society. He is also past President of the Royal Society of Chemistry.
Prof. Enemark was already well recognized for his work on metal nitrosyls
and related systems when he began to exploit the tris-pyrazolylborate ligand
as a scaffold on which to construct and study MoO2 and MoO complexes.
This work led to the synthesis and characterization of the first model that
fully mimicked the catalytic cycle of oxotransferase enzymes such as sulfite
oxidase. Enemark also played an instrumental role in the work that led to
the first crystal structure of sulfite oxidase. Since that time, Enemark has
pioneered the application of pulsed EPR methods to molybdenum enzymes
and synthetic models of their active sites; work that has led to a deep under-
standing of not simply the physical but also the electronic structures of these
systems. With over 250 publications and 10 000 citations, he is Regents Pro-
fessor of Chemistry at the University of Arizona, a former Fulbright Scholar
and recipient of the Humboldt Research Prize, among other national and
international recognitions.
Contents
Chapter 1 Spectroscopic and Electronic Structure Studies

Probing Mechanism: Introduction and Overview 1
Martin L. Kirk
1.1 I ntroduction 1
1.2 Overview 2
1.2.1 Pyranopterin Molybdenum Enzymes 2
1.2.2 Nitrogenase 8
1.3 Summary 9
Acknowledgements 10
References 10
Chapter 2 Spectroscopic and Electronic Structure Studies of

Mo Model Compounds and Enzymes 13
Martin L. Kirk
2.1 I ntroduction and Scope 13

2.2 The Pyranopterin Dithiolene and the Molybdenum
Cofactor (Moco) 16
2.2.1 General Background 16
2.2.2 Model Studies Defining the Mo–Dithiolene
Interaction in Moco 17
2.2.3 Conformational Studies of the PDT 23
2.2.4 Spectroscopic Studies of the PDT 24
2.3 Sulfite Oxidase 26
2.3.1 Active Site Structure and General Reaction
Catalyzed 26
2.3.2 Select Spectroscopic Studies of Model Systems 28

ix
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x Contents
2.3.3 S pectroscopic Studies of SO and SO-Type
Enzymes 31
2.3.4 Active Site Electronic Structure
Contributions to Reactivity 33
2.4 Xanthine Oxidoreductase (XOR) 35

Catalyzed 35
2.4.3 Spectroscopic Studies of XOR 38
2.5 Carbon Monoxide Dehydrogenase 41
Catalyzed 41
2.5.2 EPR Spectroscopic Studies of a Key
Model System 42
2.5.3 Spectroscopic Studies of CODH 43
2.6 Dimethylsulfoxide (DMSO) Reductase 47
Catalyzed 47
2.6.3 Spectroscopic Studies of DMSOR 50
2.7 MOSC Family Enzymes 53
Catalyzed 53
2.7.2 Spectroscopic Studies of MOSC Proteins 55
2.8 Perspective 58
Acknowledgements 59
References 59
Chapter 3 Electron Paramagnetic Resonance Studies of

Molybdenum Enzymes 68
Stéphane Grimaldi, Frédéric Biaso, Bénédicte Burlat,
and Bruno Guigliarelli
3.1 I ntroduction 68
3.2 Principles of EPR Techniques and Application to
Mo/ W Enzymes 69
3.2.1 Basis of EPR Spectroscopy 69
3.2.2 EPR Properties of Mo and W Enzymes 72
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Contents xi
3.3 g -Tensor Analysis for Mo/W Enzymes 75
3.3.1 g-Tensor for a d1 Configuration 75
3.3.2 Magneto-Structural Correlations in the
Mo-Enzyme Family 76
3.3.3 g-Tensor Analysis of Mo-bisPGD Active Site:

Influence of the Protein Ligands 78
3.3.4 The g-Tensor of Mo(v)-monoPPT: The
Case Study of the Sulfite Oxidase Family 81
3.3.5 g-Tensor and Substrate Binding: The Xanthine
Oxidase Family 82
3.3.6 g-Tensor Calculation: Ab-initio and DFT
Methods 83
3.3.7 g-Tensor of W(v) Species in Tungsten
Enzymes 85
3.4 Detection and Analysis of Hyperfine Couplings to
Mo/W(v) Species 86
3.4.1 Hyperfine Coupling to the Metal Ion 86
3.4.2 Superhyperfine Couplings to the Mo(v)
Species in Xanthine Oxidase Enzyme Family 90
3.4.3 Superhyperfine Couplings to Mo(v) Species
Formed in Sulfite Oxidase Enzyme Family 96
3.4.4 Superhyperfine Couplings to the Mo(v)
Species in Mo/W-bisPGD Enzymes 104
3.5 Detection and Analysis of Spin–Spin Interactions
between the Mo Cofactor and other Metal Centres 108
3.6 Concluding Remarks 110
Acknowledgements 112
References 112
Chapter 4 X-Ray Absorption Spectroscopy of Molybdenum and

Tungsten Enzymes 121
Graham N. George

4.2 The Physical Basis of X-Ray Absorption
Spectroscopy 122
4.2.1 The EXAFS 125
4.2.2 The Fourier Transform 128
4.2.3 Determination of Structural Parameters
from the EXAFS 129
4.2.4 Confusion of EXAFS Backscatterers 134
4.2.5 EXAFS Cancellation 134
4.2.6 Multiple Scattering 134
4.2.7 The EXAFS Resolution and the
Debye–Waller Term 134
4.2.8 Number of Independent Variables 136
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xii Contents
4.3 E xperimental Aspects of XAS 137
4.3.1 Sample Preparation 140
4.3.2 Data Acquisition Strategies 142
4.3.3 Fluorescence Self-Absorption Effects 143
4.3.4 Combining XAS with Other

Methods – A Holistic Approach 144
4.4 The DMSO Reductase Family of Mo and W
Enzymes 145
4.4.1 DMSO Reductase 145
4.4.2 Arsenite Oxidase 149
4.4.3 The Archaeal Tungsten Enzymes 150
4.4.4 Other DMSO Reductase Family
Members 151
4.5 The Xanthine Oxidase Family of Mo Enzymes 152
4.5.1 Xanthine Oxidase 152
4.5.2 Carbon Monoxide Dehydrogenase 153
4.6 The Sulfite Oxidase Family of Mo Enzymes 154
4.7 Nitrogenase 158
4.8 Concluding Remarks 161
References 162
Chapter 5 Electrochemistry of Molybdenum and Tungsten

Enzymes 168
Palraj Kalimuthu and Paul V. Bernhardt

5.1.1 The Mo and W Enzyme Families 168
5.1.2 Enzyme Electrochemistry 169
5.2 Xanthine Oxidase Family 172
5.2.1 Xanthine Oxidoreductase 172
5.2.2 Aldehyde Oxidoreductase 180
5.3 Sulfite Oxidase Family 181
5.3.1 Sulfite Oxidoreductase 182
5.3.2 Eukaryotic Nitrate Reductase 192
5.4 DMSO Reductase Family 196
5.4.1 DMSO Reductase 197
5.4.2 DMS Dehydrogenase 203
5.4.3 Bacterial Nitrate Reductase 203
5.4.4 Arsenite Oxidase 208
5.4.5 Ethylbenzene Dehydrogenase 211
5.4.6 Formate Dehydrogenase 212
5.4.7 Glyceraldehyde 3-Phosphate
Oxidoreductase 214
5.5 Conclusions 214
Acknowledgement 215
References 215
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Contents xiii
Chapter 6 Nitrogen Fixation in Nitrogenase and Related
Small-Molecule Models: Results of DFT Calculations 223
Felix Tuczek

6.1.1 Structure and Function of Nitrogenase 224
6.1.2 Fe-Protein Cycle 224
6.1.3 Metal Clusters Within the MoFe-Protein 226
6.1.4 Thorneley–Lowe Cycle 227
6.1.5 Site-Directed Mutagenesis Experiments 227
6.1.6 Trapping and Spectroscopic Characterization
of Intermediates of N2 Reduction: Towards
an Experimentally Derived Mechanism of
Nitrogenase 228
6.2 DFT Treatments of N2 Reduction in Model
Systems 229
6.2.1 Schrock Cycle 231
6.2.2 Nishibayashi’s System 238
6.2.3 Chatt Cycle 239
6.2.4 Reduction and Protonation of N2 at Cubane
Clusters 241
6.2.5 Reduction and Protonation of N2 at Iron
Complexes 242
6.3 DFT Calculations on the FeMoco and its
Reactivity with N2 246
6.3.1 Noodleman and Coworkers 246
6.3.2 Nørskov and Coworkers 252
6.3.3 Blöchl, Kästner et al 256
6.3.4 Dance 259
6.3.5 Further Theoretical Studies 261
6.3.6 Mo(iii) Charge State of FeMoco 264
6.4 Summary and Conclusions 266
Acknowledgement 266
References 267
Chapter 7 Computational Studies of Molybdenum and Tungsten

Enzymes 275
Ulf Ryde, Geng Dong, Jilai Li, Milica Feldt, and
Ricardo A. Mata

7.2 Computational Methods to Study Metalloenzymes 279
7.2.1 QM Methods 280
7.2.2 Hybrid QM/QM Calculations 283
7.2.3 QM-Cluster Calculations 283
7.2.4 QM/MM Calculations 284
7.2.5 How to Model the MPT Ligand 287
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xiv Contents
7.3 DMSO Reductase 289
7.4 Sulfite Oxidase 298
7.5 Xanthine Oxidase 306
7.6 Comparison of the Three Families 311
7.7 Conclusions 314

References 316
Subject Index 322

Published on 30 September 2016 on http://pubs.rsc.org | doi:10.1039/9781782628842-00001
Chapter 1
Spectroscopic and Electronic

Structure Studies Probing
Mechanism: Introduction and
Overview
Martin L. Kirka
a
Department of Chemistry and Chemical Biology, The University of New
Mexico, MSC03 2060, 1 University of New Mexico, Albuquerque,
New Mexico 87131-0001, USA
*E-mail: mkirk@unm.edu
1.1 Introduction
The first Gordon Research Conference on “Molybdenum and Tungsten
Enzymes” was held July 4–9 1999 at Plymouth State College (now Plym-
outh State University) in New Hampshire and was chaired by Ed Stiefel
and Russ Hille. This meeting proved to be a transformative one for our
field in that it provided an intellectual forum for molecular biologists, syn-
thetic chemists, enzymologists, theorists, crystallographers and spectros-
copists to converge, discuss their latest results and develop long-standing
relationships that would foster new collaborations that have allowed us to
understand the structure and function of Mo- and W-containing enzymes
as well as the intricate details of their catalytic mechanisms of activity.

1
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2 Chapter 1
These enzymes continue to be the subject of intense research efforts,
and this is a direct result of their unusual geometric and electronic struc-
tures, their key roles in the global C, N and S cycles, their pharmacologi-
cal importance, and their importance in human health. This volume will
detail how spectroscopy, structure, electrochemistry and theory have been

used to develop a comprehensive description of the active site electronic
structure contributions to reactivity in pyranopterin Mo enzymes and the
Mo-dependent nitrogenase. A particular emphasis is placed on how these
important studies have been used to reveal critical components of enzyme
mechanisms.
1.2 Overview
1.2.1 Pyranopterin Molybdenum Enzymes
With the sole exception of nitrogenase, the pyranopterin Mo (and W) enzymes
are the only metalloenzymes that utilize second-1,2 (and third-) row transition
metal ions to catalyze a myriad of redox transformations involving enzyme
substrates.3,4 The uniqueness of these enzymes is further underscored by the
fact that they possess the molybdenum cofactor (Moco, Figure 1.1),5,6 which
is comprised of a high-valent MoIV,V,VI (WIV,V,VI) ion coordinated by a unique
dithiolene ligand (the pyranopterin dithiolene). This dithiolene ligand is
connected to a pterin ring system by a pyran ring that may be in either a
closed (typical) or an open ring configuration. The pyranopterin dithiolene
(PDT) is a highly complex ligand (vide infra) that is unique to the mononu-
clear Mo (W) enzymes. The remarkable nature of this ligand is exemplified
by its potential for electronic flexibility, including changing its redox and/or
tautomeric state to exert additional control of the Mo redox potential.6 The
pyranopterin molybdenum enzymes have been historically divided into three
broad families: the sulfite oxidase (SO) family, the xanthine oxidase (XO) fam-
ily and the dimethylsulfoxide reductase (DMSOR) family of enzymes (Figure
1.2).5 This broad classification has been based on the coordination geometry
of their active sites, the nature of their respective protein folds and the type
and breadth of reactions that are catalyzed by the enzymes.
Figure 1.1
The molybdenum cofactor (Moco), comprised of a Mo ion bound to a
pyranopterin dithiolene (PDT) chelate. Note that the PDT shown here is
in the fully reduced “tetrahydro” oxidation state.5,6
View Online
Spectroscopic and Electronic Structure Studies Probing Mechanism 3

The vast majority of pyranopterin Mo (W) enzymes catalyze the formal trans-
fer of an oxygen atom between the substrate and the Mo ion. The generalized
oxygen atom transfer reactions possess the following stoichiometry:5,7–9
E-MoVI + R + H2O → E-MoIV + R–O + 2H+

Regarding substrate oxidations, reductive oxygen atom transfer is given

by the reverse of this reaction. In marked contrast to the oxotransferases,
substrate hydroxylations (formal insertion of an O atom into a substrate C–H
bond) follow the reaction stoichiometry given below:
E-MoVI + R–H + H2O → E-MoIV + R–OH + 2H+

Excellent reviews3,5,7–17 have highlighted recent advances in our under-
standing of these enzymes, in addition to reviews that have covered the
Mo-dependent nitrogenase,18–20 which will be introduced later in this chapter.
More specific reviews have focused on the sulfite oxidase,21,22 DMSO reduc-
tase23 and xanthine oxidase7,24 enzyme families, with additional reviews cov-
ering Moco biosynthesis,25–27 spectroscopic studies17,28 and computational
probes of the enzyme reaction coordinates.16,28
Although it has long been suggested that the PDT may play key roles in facil-
itating vectorial electron transfer, modulating enzyme reduction potentials
and providing an anchor for the Mo ion in the catalytic active site, there is still
much to be learned about this complex ligand. Some of the latest work on the
PDT has detailed a relationship between pyranopterin dithiolene geometric
structure and enzyme function.6 A bioinformatics study of 309 pyranopterins
from 109 separate enzyme structures showed that these enzymes possessed
Figure 1.2
Top: bond line drawings for the oxidized and reduced members of the
three canonical pyranopterin Mo enzyme families (SO, DMSOR and
XO). Bottom: active site coordination geometries for SO, DMSOR, and
XO as determined by X-ray crystallography.
View Online
4 Chapter 1
geometries that can be described by a well-defined distortion coordinate that
is related to the nature of PDT out-of-plane distortions that were revealed by
X-ray crystallography.6 This distortion coordinate was analyzed in the context of
DFT calculations that were performed on geometry-optimized PDT structures.
The results suggest that differences in the nature of the PDT out-of-plane dis-
tortions are related to PDTs that adopt different oxidation states. Specifically,
the analysis suggests that biological PDTs may not all exist in the fully reduced
“tetrahydro” oxidation state, as has been previously thought.5 These research-
ers hypothesized that the PDT may also be present in two electron oxidized
“dihydro” forms.6 Remarkably, the observed PDT distortions can be associated
with specific enzyme families. For example, the PDT distortions observed for
XO family enzymes are consistent with the PDT being in the fully reduced “tet-
rahydro” oxidation state, while SO family PDTs adopt “dihydro” structures.
Interestingly, DMSOR family enzymes, which contain two PDTs coordinated
to Mo, appear to possess one PDT that displays an SO type distortion and one
PDT possessing an XO type distortion. The results are of interest in that they
suggest a link between enzyme function and the oxidation state(s) of the PDT.6
In support of this idea, the first structurally characterized oxomolybdenum
complex to incorporate a pyranopterin dithiolene ligand was found to possess
the ligand in the “dihydro” oxidation state.29 A combination of X-ray crystal-
lography and 1H NMR spectroscopy was used to show that this complex pos-
sessed a complete pyranopterin dithiolene ligand, and that reversible pyran
ring opening and closing may represent a dynamic process in pyranopterin
molybdenum enzymes. Although these addressed apparent relationships
between PDT geometric structure, oxidation state and enzyme function, our
understanding of how PDT electronic structure contributes to enzyme catalysis
remains to be determined.
From a spectroscopic and electronic structure viewpoint, the pyranopterin
molybdenum (and tungsten) containing enzymes are unique among metal-
loenzymes. This primarily derives from the terminal Mo-oxo ligation cou-
pled with the high oxidation states accessible to the metal ion during the
course of catalysis. This results in a large splitting of the t2g orbital set with
the Mo(xy) redox active orbital being well separated energetically from the
Mo–Ooxo dπ* antibonding orbitals.30 Thus, the Mo(v) ion possesses an (xy)1
configuration with the redox orbital oriented perpendicular to a Mo–Ooxo
vector30 and the Mo(iv) ion possesses a low-spin (xy)2 configuration with a
diamagnetic ground state. Spectroscopic studies probing the Mo ion in pyra-
nopterin Mo enzymes have not been trivial. This results from the fact that
the majority of pyranopterin Mo enzymes possess strongly absorbing flavin,
iron–sulfur and/or heme chromophores5 that mask the electronic absorp-
tion spectra associated with the Mo active sites. Furthermore, this problem
is exacerbated by the fact that the only relevant paramagnetic state is the
Mo(v) oxidation state, which represents an obligatory catalytic intermedi-
ate in the electron transfer regeneration half-reaction that progresses via
sequential one-electron transfers that interconvert the diamagnetic Mo(iv)
and Mo(vi) oxidation states. Thus, in order to use high resolution paramag-
netic probes of pyranopterin Mo enzyme active site electronic structure, it is
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31
necessary to either trap reaction intermediates or study inhibited enzyme
forms.32,33 This critical issue cannot be overstated, as the general inability to
probe resting, fully oxidized or fully reduced enzyme forms using paramag-
netic spectroscopies has provided a significant challenge to understanding
crucial interrelationships between spectroscopic features of the active sites

and their corresponding geometric and electronic structures.
These caveats aside, paramagnetic spectroscopies, including EPR, ESEEM,
ENDOR, MCD and Mössbauer, have played critical roles in determining the
electronic structure of the Mo(v) state and have contributed to a greater
understanding of pyranopterin molybdenum enzyme reaction coordinates.
The first spectroscopic studies of pyranopterin Mo enzyme geometric and
electronic structure employed EPR spectroscopy. Notable among these are
the early work of Bray and coworkers, who used EPR to investigate rapidly
appearing Mo(v) EPR signals in reduced forms of XO.34 These early EPR
studies were often calibrated against data collected on small molecule mod-
els,35,36 as there were not yet any high-resolution protein crystal structures.
The xanthine oxidase very rapid species,37,38 which can be trapped under
turnover conditions with specific substrates,39 represents a clear example of
how multiple spectroscopic probes of an enzyme intermediate have played
a significant role in enhancing our understanding of the enzyme's mecha-
nism. The very rapid intermediate is a Mo(v)-product species that has been
extensively probed by a combination of EPR37–39 (Figure 1.3), ENDOR40 and
MCD31 spectroscopies. Collectively, these data have been used to show that
Figure 1.3
X-band EPR spectrum of the xanthine oxidase very rapid intermediate
generated with 2-hydroxy-6-methylpurine as reducing substrate. Data
were acquired at 150 K, 9.47 GHz and 10 mW microwave power. Note
the high value for g1, which has been used in conjunction with 33S
hyperfine analysis to indicate the presence of a highly covalent Mo=S
π bonding scheme in very rapid and, by inference, the oxidized Mo(vi)
form of the enzyme. Adapted with permission from ref. 31. Copyright
(1999) American Chemical Society.
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6 Chapter 1
the very rapid intermediate (1) possesses an apical oxo and an equatorial
sulfido ligand that are oriented cis relative to one another,31 (2) possesses
a highly covalent Mo=S d–p π bonding interaction, (3) does not involve the
formation of an organometallic Mo–C bond in the catalytic cycle of the
enzyme40 and (4) possesses product bound to the Mo ion as the enolate tau-
tomer.40 The combination of computational, spectroscopic and reactivity
studies on xanthine oxidase very rapid has contributed greatly to our under-
standing of the reductive half-reaction in xanthine oxidase and related Mo
hydroxylase family enzymes.3 Of course, the xanthine oxidase very rapid
story is only one of many studies where paramagnetic spectroscopies have
proven to be crucial for developing a greater understanding of pyranopterin
molybdenum enzyme electronic structure contributions to reactivity.3,24
Diamagnetic probes of enzyme electronic structure have primarily involved
electronic absorption and resonance Raman spectroscopies. It is import-
ant to note that when spectroscopic studies on the enzymes can be directly
compared to analogous studies on small molecule analogs of their active
sites, tremendous insight into the relationships between electronic and geo-
metric structure contributions begins to emerge. This is due, in part, to the
fact that the small molecule analogs do not possess the competing chromo-
phores that are present in many of the enzymes, and they allow for specific
structural components found in the enzymes to be evaluated individually
(Figure 1.4). This has resulted in the detailed spectral probing of numer-
ous elegantly designed small molecules using electronic absorption spec-
troscopy, S K-edge XAS, resonance Raman and MCD spectroscopies.30,41–59
Spectroscopic studies on diamagnetic model compounds in the catalytically
relevant Mo(iv) and Mo(vi) oxidation states have also contributed greatly to
Figure 1.4
Specific structural components that are found in pyranopterin Mo
enzymes, which have also been incorporated into small-molecule
model systems for detailed spectroscopic studies.
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our understanding of pyranopterin Mo enzymes in these “spectroscopically
challenged” oxidation states.
The initial contribution to this volume provides a detailed overview of
how spectroscopy and computations have been used in concert to probe the
canonical members of each pyranopterin Mo enzyme family, as well as the

pyranopterin dithiolene ligand itself. The discussion focuses on how a com-
bination of enzyme geometric structure, spectroscopy and biochemical data
have been used to arrive at an understanding of electronic structure contri-
butions to reactivity in all of the major pyranopterin Mo enzyme families.
A unique aspect of this discussion is that spectroscopic studies on relevant
small molecule model compounds have been melded with analogous studies
on the enzyme systems to arrive at a sophisticated description of active site
electronic structure. As the field moves forward, it will become increasingly
important to understand the structure, function and reaction mechanisms
for the numerous non-canonical (i.e. beyond sulfite oxidase, xanthine oxidase,
DMSO reductase) pyranopterin Mo enzymes.
Guigliarelli and coworkers then present a detailed overview of how EPR
spectroscopy has been applied to the study of pyranopterin Mo enzymes.
A specific emphasis has been placed on the challenging nature of studying
reaction intermediates and how analysis of the magnitude and anisotropy
of the g-tensor have contributed to a greater understanding of the enzymes.
They discuss advances in pulsed EPR techniques, including ENDOR, ESEEM
and HYSCORE, and how these have been used with isotope perturbation and
computational studies to understand the nature of the hyperfine tensor and
how this relates to structure and mechanism. Finally, they detail how one
may obtain long-range structural details from spin–spin coupling between
distinct paramagnetic centers in the enzymes.
The first pyranopterin molybdenum enzyme to be fully characterized by
X-ray crystallography was the aldehyde oxidoreductase from Desulfovibrio
gigas.60 The subsequent determination of numerous molybdenum protein
structures, including nitrogenase, by X-ray crystallography have revolution-
ized the way we think about geometric structure contributions to catalysis.
These data have provided electronic structure researchers, synthetic chem-
ists and spectroscopists with a structural reference for the starting point in
the catalytic cycles of these enzymes. When combined with EXAFS analysis, a
clear picture of the active site metal–ligand first coordination sphere begins
to emerge for both the oxidized and reduced states of the enzyme. George
has provided a thorough tutorial-type presentation of X-ray absorption spec-
troscopy (XAS), including the analysis of EXAFS data, before entering into
a discussion of how XAS has enhanced our understanding of pyranopterin
Mo and W enzymes. A thoughtful component of the contribution details the
relative strengths and weaknesses of XAS as a structural probe by using a
series of illustrative examples.
As mentioned previously, the vast majority of pyranopterin Mo enzymes
catalyze two-electron redox reactions that are coupled to the formal trans-
fer of an oxygen atom between the Mo center and the substrate. As such,
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8 Chapter 1
electrochemical studies of the enzymes provide insight into these two-electron
redox processes, in addition to the one-electron transfer events that are asso-
ciated with the completion of the catalytic cycle. Kalimuthu and Bernhardt
have provided a wonderful and timely discussion of direct and mediated pro-
tein electrochemistry of pyranopterin Mo enzymes that begins with a tutorial

on enzyme electrochemistry (DC voltammetry). DC voltammetry is of particu-
lar utility as it is well suited for probing metalloenzymes that are undergoing
catalysis (i.e. oxidizing or reducing substrates) since the observed current (cat-
alytic current) is amplified. A description of how XOR-, sulfite oxidase-, nitrate
reductase- and DMSO reductase-based electrochemistry can be used for the
development of amperometric biosensors is included in their contribution.
Theory has played a central role in our understanding of molybdenum
enzyme active site structure, dynamics, mechanism, spectroscopy and elec-
tronic structure contributions to reactivity. Ryde and coworkers have reviewed
how computations have enhanced our understanding of enzymes that belong
to the SO, DMSOR and XOR enzyme families. Particular emphasis is given
to understanding the effects of basis set size, nature of the functional, solva-
tion and dispersion effects, etc. on the determination of computed reaction
coordinate energies, including transition states. The quantum mechanical
approaches that are highlighted have been diverse. Modern DFT and config-
uration interaction methodologies will continue to be utilized to understand
these remarkable biological catalysts that employ a second-row transition
metal ion. Coupled with the ever-increasing power of modern computers, we
can expect continued and accelerated applications of theory to provide addi-
tional insight into the interpretation of experimental results, and to greatly
assist in solving some of the most difficult problems related to these enzymes.
1.2.2 Nitrogenase
The Mo dependent nitrogenase is the sole exception among all of the Mo con-
taining enzymes in that it does not possess a pyranopterin dithiolene ligand
bound to the Mo ion. Molybdenum-dependent nitrogenase consists of an Fe
protein and a MoFe protein, the latter of which possesses an 8Fe7S P-cluster
and an Fe7S9Mo cluster possessing a homocitrate bound to Mo and an
unusual carbide ligand embedded in the center of the core. The FeMo-cofactor
(FeMo-co) represents the locus of dinitrogen reduction to ammonia according to:
N2 + 8e− + 16MgATP + 8H+ → 2NH3 + H2 + 16MgADP + 16Pi

N2 + 6e− + 6H+ → 2NH3
The catalytic mechanism of nitrogenase is discussed in terms of the Lowe–

Thorneley scheme (Figure 1.5), which details eight sequential proton coupled
electron transfer steps in the catalytic cycle. The importance of nitrogenase
is underscored by its dominant role in biogeochemical nitrogen fixation.
Spectroscopic studies probing the Mo ion in nitrogenase have also not been
trivial due to the large protein iron content and the complex nature of the
View Online

Figure 1.5 modified Lowe–Thorneley scheme for nitrogenase. Adapted from

A
ref. 18.
exchange coupled paramagnetic spin centers. For nitrogenase, Mössbauer

spectroscopy has been used to probe iron oxidation states and spin states
of FeMoco, and has proven to be complementary to the high information
content of paramagnetic resonance spectroscopies. The goal of these spec-
troscopic studies is to provide a geometric and electronic structure descrip-
tion of the various coupled electron–proton transfer events that make up the
various components of the Lowe–Thorneley scheme in order to develop a
detailed understanding of the nitrogenase catalyzed six-electron reduction
of dinitrogen to ammonia. Nitrogenase provides a prime example of how
spectroscopic studies can be used to gain insight into enzyme structure and
mechanism. Most recently, pulsed EPR and ENDOR methods have been cou-
pled with approaches that include freeze-quench and the study of enzyme
variants in order to identify several catalytic intermediates and their relation-
ship to the nitrogenase catalytic cycle.
As such, the final chapter of the volume effectively ends as it begins, with
a detailed discussion of how model and enzyme structural, computational
and spectroscopic studies have led to an increased understanding of enzyme
(nitrogenase) reactivity. Here, Tuczek provides an excellent state-of-the-art
review of how these approaches have been utilized in order to unravel the
intricacies of how Nature fixes nitrogen. He begins with a description of how
model studies have provided detailed information regarding how dinitro-
gen binds to metal centers and is subsequently activated for cleavage of the
strong N≡N bond by a series of protonation steps and the introduction of
reducing equivalents. This work is evaluated in the context of computational
studies on the enzyme system, with a specific emphasis on correlating the
computational work to available experimental data, including spectroscopic
studies on nitrogenase.
1.3 Summary
Clearly, there have been numerous advances in our understanding of how
pyranopterin Mo enzymes and nitrogenase function, including how the
underpinning electronic structure of their respective catalytic active sites
contributes to their unique reactivities. The individual contributions to
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10 Chapter 1
this volume highlight how we, as a community, have greatly expanded the
knowledge base by using a combined structural, spectroscopic, voltam-
metric and computational approach to ultimately understand the nature
of their catalytic cycles. Of course, this would not be possible without the
numerous contributions from researchers engaged in the biological, struc-

tural and small-molecule studies that have been detailed in the two other
volumes.
Acknowledgements
M. L. K. would like to thank all of his graduate students, postdoctoral associ-
ates and collaborators who have contributed to the various works that have
been described in this chapter. M. L. K. also acknowledges the National Insti-
tutes of Health (GM-057378) for continued support of the author’s work, part
of which is included in this chapter.
References
1. J. J. R. Frausto da Silva and R. J. P. Williams, The Biological Chemistry of the
Elements: The Inorganic Chemistry of Life, Clarendon Press, Oxford, 1991.
2. S. Lippard and J. Berg, Principles of Bioinorganic Chemistry, University
Science Books, Mill Valley, 1994.
3. M. L. Kirk and B. Stein, in Comprehensive Inorganic Chemistry II (Second
Edition), ed. R. Jan and P. Kenneth, Elsevier, Amsterdam, 2013, p. 263.
4. R. Hille, J. Hall and P. Basu, Chem. Rev., 2014, 114, 3963.
5. R. Hille, Chem. Rev., 1996, 96, 2757.
6. R. A. Rothery, B. Stein, M. Solomonson, M. L. Kirk and J. H. Weiner, Proc.
Natl. Acad. Sci. U. S. A., 2012, 109, 14773.
7. R. Hille, Arch. Biochem. Biophys., 2005, 433, 107.
8. R. Hille, J. Retey, U. Bartlewski-Hof, W. Reichenbecher and B. Schink,
FEMS Microbiol. Rev., 1998, 22, 489.
9. R. Hille, JBIC, J. Biol. Inorg. Chem., 1996, 1, 397.
10. L. Noodleman, T. Lovell, T. Q. Liu, F. Himo and R. A. Torres, Curr. Opin.
Chem. Biol., 2002, 6, 259.
11. R. Hille, JBIC, J. Biol. Inorg. Chem., 1998, 3, 559.
12. R. Hille, Molybdenum enzymes containing the pyranopterin cofactor: An
overview, Marcel Dekker, Inc., New York, 2002, vol. 39.
13. R. Hille, T. Nishino and F. Bittner, Coord. Chem. Rev., 2011, 255, 1179.
14. H. Sugimoto and H. Tsukube, Chem. Soc. Rev., 2008, 37, 2609.
15. A. Majumdar and S. Sarkar, Coord. Chem. Rev., 2011, 255, 1039.
16. S. Metz and W. Thiel, Coord. Chem. Rev., 2011, 255, 1085.
17. M. J. Pushie and G. N. George, Coord. Chem. Rev., 2011, 255, 1055.
18. B. M. Hoffman, D. Lukoyanov, D. R. Dean and L. C. Seefeldt, Acc. Chem.
Res., 2013, 46, 587.
19. B. M. Hoffman, D. R. Dean and L. C. Seefeldt, Acc. Chem. Res., 2009, 42,
609.
View Online

20. P. C. Dos Santos, R. Y. Igarashi, H.-I. Lee, B. M. Hoffman, L. C. Seefeldt
and D. R. Dean, Acc. Chem. Res., 2005, 38, 208.
21. C. Feng, G. Tollin and J. H. Enemark, Biochim. Biophys. Acta, Proteins
Proteomics, 2007, 1774, 527.
22. G. J. Workun, K. Moquin, R. A. Rothery and J. H. Weiner, Microbiol. Mol.

Biol. Rev., 2008, 72, 228.
23. A. G. McEwan and U. Kappler, Aust. Biochemist, 2004, 35, 17.
24. B. Stein and M. Kirk, JBIC, J. Biol. Inorg. Chem., 2015, 20, 183.
25. R. R. Mendel and G. Schwarz, Coord. Chem. Rev., 2011, 255, 1145.
26. G. Schwarz and R. R. Mendel, Annu. Rev. Plant Biol., 2006, 57, 623.
27. P. Basu and S. J. N. Burgmayer, Coord. Chem. Rev., 2011, 255, 1016.
28. M. L. Kirk, S. Knottenbelt and A. Habtegabre, in Computational Inorganic
and Bioinorganic Chemistry, ed. E. I. Solomon, R. A. Scott and B. R. King,
Wiley, 2009, p. 614.
29. B. R. Williams, Y. C. Fu, G. P. A. Yap and S. J. N. Burgmayer, J. Am. Chem.
Soc., 2012, 134, 19584.
30. F. E. Inscore, R. McNaughton, B. L. Westcott, M. E. Helton, R. Jones, I. K.
Dhawan, J. H. Enemark and M. L. Kirk, Inorg. Chem., 1999, 38, 1401.
31. R. M. Jones, F. E. Inscore, R. Hille and M. L. Kirk, Inorg. Chem., 1999, 38,
4963.
32. M. Shanmugam, B. Zhang, R. L. McNaughton, R. A. Kinney, R. Hille and
B. M. Hoffman, J. Am. Chem. Soc., 2010, 132, 14015.
33. J. Sempombe, B. Stein and M. L. Kirk, Inorg. Chem., 2011, 50, 10919.
34. R. C. Bray and T. Vänngård, Biochem. J., 1969, 114, 725.
35. R. Greenwood, G. Wilson, J. Pilbrow and A. Wedd, J. Am. Chem. Soc.,
1993, 115, 5385.
36. G. L. Wilson, R. J. Greenwood, J. R. Pilbrow, J. T. Spence and A. G. Wedd,
J. Am. Chem. Soc., 1991, 113, 6803.
37. S. Gutteridge and R. C. Bray, Biochem. J., 1980, 189, 615.
38. R. Bray, S. Gutteridge, D. Stotter and S. Tanner, Biochem. J., 1979, 177,
357.
39. R. McWhirter and R. Hille, J. Biol. Chem., 1991, 266, 23724.
40. P. Manikandan, E. Choi, R. Hille and B. Hoffman, J. Am. Chem. Soc., 2001,
123, 2658.
41. H. Sugimoto, H. Tano, K. Suyama, T. Kobayashi, H. Miyake, S. Itoh, R. P.
Mtei and M. L. Kirk, Dalton Trans., 2011, 40, 1119.
42. K. G. Matz, R. P. Mtei, R. Rothstein, M. L. Kirk and S. J. N. Burgmayer,
Inorg. Chem., 2011, 50, 9804.
43. H. Sugimoto, S. Tatemoto, K. Suyama, H. Miyake, R. P. Mtei, S. Itoh and
M. L. Kirk, Inorg. Chem., 2010, 49, 5368.
44. K. G. Matz, R. P. Mtei, B. Leung, S. J. N. Burgmayer and M. L. Kirk, J. Am.
Chem. Soc., 2010, 132, 7830.
45. R. L. McNaughton, B. S. Lim, S. Z. Knottenbelt, R. H. Holm and M. L.
Kirk, J. Am. Chem. Soc., 2008, 130, 4628.
46. C. J. Doonan, N. D. Rubie, K. Peariso, H. H. Harris, S. Z. Knottenbelt, G.
N. George, C. G. Young and M. L. Kirk, J. Am. Chem. Soc., 2008, 130, 55.
View Online
12 Chapter 1
47. K. Peariso, M. E. Helton, E. N. Duesler, S. E. Shadle and M. L. Kirk, Inorg.
Chem., 2007, 46, 1259.
48. S. J. N. Burgmayer, M. Kim, R. Petit, A. Rothkopf, A. Kim, S. BelHam-
dounia, Y. Hou, A. Somogyi, D. Habel-Rodriguez, A. Williams and M. L.
Kirk, J. Inorg. Biochem., 2007, 101, 1601.

49. F. E. Inscore, S. Z. Knottenbelt, N. D. Rubie, H. K. Joshi, M. L. Kirk and J.
H. Enemark, Inorg. Chem., 2006, 45, 967.
50. R. L. McNaughton, S. Mondal, V. N. Nemykin, P. Basu and M. L. Kirk,
Inorg. Chem., 2005, 44, 8216.
51. R. L. McNaughton, M. E. Helton, M. M. Cosper, J. H. Enemark and M. L.
Kirk, Inorg. Chem., 2004, 43, 1625.
52. N. D. Rubie, P. Katrina, C. Doonan, G. N. George, C. G. Young and M. L.
Kirk, J. Am. Chem. Soc., 2002, 224, U642.
53. K. Peariso, R. L. McNaughton and M. L. Kirk, J. Am. Chem. Soc., 2002,
124, 9006.
54. V. N. Nemykin, S. R. Davie, S. Mondal, N. Rubie, M. L. Kirk, A. Somogyi
and P. Basu, J. Am. Chem. Soc., 2002, 124, 756.
55. S. R. Davie, N. D. Rubie, B. S. Hammes, C. J. Carrano, M. L. Kirk and P.
Basu, Inorg. Chem., 2001, 40, 2632.
56. R. L. McNaughton, A. A. Tipton, R. R. Conry and M. L. Kirk, J. Am. Chem.
Soc., 2000, 219, U782.
57. M. L. Kirk, M. E. Helton, N. E. Gruhn and R. L. McNaughton, J. Am. Chem.
Soc., 2000, 219, U782.
58. M. Helton, N. Gruhn, R. McNaughton and M. Kirk, Inorg. Chem., 2000,
39, 2273.
59. F. Inscore, R. McNaughton, B. Westcott, M. Helton, R. Jones, I. Dhawan,
J. Enemark and M. Kirk, Inorg. Chem., 1999, 38, 1401.
60. M. J. Romão, M. Archer, I. Moura, J. J. G. Moura, J. LeGall, R. Engh, M.
Schneider, P. Hof and R. Huber, Science, 1995, 270, 1170.
Chapter 2
Spectroscopic and Electronic

Structure Studies of Mo Model
Compounds and Enzymes
Martin L. Kirka
a
Department of Chemistry and Chemical Biology, The University of New
Mexico, MSC03 2060, 1 University of New Mexico, Albuquerque, New Mexico
87131-0001, USA
*E-mail: mkirk@unm.edu
2.1 Introduction and Scope

The pyranopterin molybdenum (Mo) enzymes are unique in that they, with
the exception of nitrogenase, are the only metalloenzymes that utilize the
molybdenum ion as the critical constituent of their active sites.1–4 Addition-
ally, these enzymes possess a special ligand, the pyranopterin dithiolene
(PDT), which when bound to molybdenum forms what is referred to as the
molybdenum cofactor (Moco) (Figure 2.1). The dithiolene component of the
PDT binds the molybdenum ion in a bidentate manner forming highly cova-
lent Mo–S bonds. The pyranopterin Mo enzymes are the subject of intense
study, as they are found in almost all life forms from archaea to humans, are
essential for human life, play critical roles in the biogeochemical C, N and S
cycles and possess an emerging pharmacological importance.

13
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14 Chapter 2
For the purpose of this work, we divide the pyranopterin molybdenum
enzymes into five groups: sulfite oxidase (SO) type enzymes, xanthine oxidore-
ductase (XOR) type enzymes, carbon monoxide dehydrogenase (CODH),
dimethylsulfoxide reductase (DMSOR) type enzymes and the molybdenum
cofactor sulfurase C-terminal (MOSC) domain proteins. Their basic oxidized

and reduced geometric structures are depicted in Figure 2.2. Although there
are exceptions, the molybdenum ion in these enzymes typically catalyzes two
electron redox reactions and redox cycles between the Mo(iv) and Mo(vi) oxi-
dation states with the paramagnetic Mo(v) state being an obligatory catalytic
intermediate in the electron transfer half-reaction of the enzymes. For most
pyranopterin Mo enzymes, the two electron chemistry that they catalyze
involves the formal transfer of an oxygen atom between the substrate and
the active site catalyst according to the general reaction:1,5–7
E-MoVI + R + H2O → E-MoIV + R–O + 2H+
Notable exceptions occur in the formate dehydrogenases and acetylene

hydratase, and possibly in bacterial YedY8–10 and the mitochondrial ami-
doxime reducing component (mARC) enzyme. A markedly different type of
reaction is catalyzed by members of the xanthine oxidoreductase (XOR) fam-
ily of enzymes and involves the formal insertion of an oxygen atom into a
substrate C–H bond. Thus, these enzymes are often categorized as hydrox-
ylases.11–15 However, the reactivity patterns of the XORs differ substantially
Figure 2.1
Left: the pyranopterin dithiolene (PDT). Right: one form of the molyb-
denum cofactor (Moco).
Figure 2.2
Bond line drawings of consensus structures for the five groups of pyra-
nopterin molybdenum enzymes discussed in this chapter. For DMSOR
family enzymes, X = H2O/OH, OSer, OAsp, SCys, SeSec, or is absent entirely.38
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Spectroscopic and Electronic Structure Studies of Mo Model 15

from the monooxygenase enzymes, which insert an oxygen atom from metal
activated dioxygen into a C–H bond and consume reducing equivalents in
the process. The XORs catalyze formal hydroxylation reactions of the type:
E-MoVI + R–H + H2O → E-MoIV + R–OH + 2H+
where the oxygen atom formally inserted into the substrate C–H bond derives
from metal activated water and reducing equivalents are generated rather
than consumed.1
Molybdenum is a second-row transition element and a strong octahedral
ligand field (ΔOh) is assured (i.e. a large t2g–eg splitting). As a result, the eg orbit-
als are never populated. However, the active site coordination geometries are
markedly lower than octahedral and this leads to a large splitting of the t2g
orbital set.16–18 This is important, since it leads to low-spin d2 electronic con-
figurations even in the absence of a terminal oxido ligand.18 To date, the high-
spin S = 1 d2 configuration for the Mo(iv) ion in pyranopterin Mo enzymes has
yet to be observed.19 Thus, the electronic structures of molybdenum sites in
proteins differ considerably from first-row transition elements that can pos-
sess high-spin electronic configurations. The active site Mo in the enzymes is
found bound to both very hard (i.e. terminal oxo) and very soft (thiol/dithi-
olene) donor ligands, contributing their mechanistic complexity and an
extreme flexibility with respect to their electronic structures. When terminal
oxo ligands are present, the orientation of the redox active molecular orbital
(RAMO) is defined by the orientation of these strong σ-donor, π-donor ligands.
For mono-oxido sites, this orbital is commonly referred to as the dxy orbital and
it is always oriented orthogonal to the terminal oxido donor. This has led to an
“oxido gate” hypothesis for pyranopterin Mo enzymes. Here, having a single
terminal oxido donor oriented orthogonal to the dithiolene chelate is a neces-
sary prerequisite for facile electron transfer regeneration of the active site.16,20
A number of excellent reviews1,5–7,21–36 have recently appeared that sum-
marize our knowledge of pyranopterin molybdenum enzymes, including the
detailed structural knowledge of these enzymes using X-ray crystallography
and XAS/EXAFS. We will not provide a detailed overview of the structures
of enzymes discussed in this work. The scope of this chapter is such that it
focuses on very select spectroscopic and electronic structure studies of pyra-
nopterin molybdenum enzymes and small molecule analogs of their active
sites, with a particular emphasis being placed on work from the author's
laboratories. We do not attempt to be comprehensive, but rather attempt to
illuminate through the use of key examples. We have attempted to highlight
specific examples of spectroscopic studies where data on both the enzymes
and models can be compared. These examples have been chosen to show
the power of a combined spectroscopic approach applied to both enzymes
and models. The spectroscopic studies are augmented by detailed electronic
structure calculations that have been calibrated to structural and spectro-
scopic data. This powerful approach can reveal exquisitely intimate details
regarding how the underpinning electronic structure of these unique metal-
loenzymes contributes to their function and reactivity patterns.
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SPECIFIC CHARACTER.
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DESCRIPTION.
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surface, pressed to the stem, and smooth, having very short foot-stalks.
The Flowers are numerous, and terminate the smaller branches in
bunches; the foot-stalks are very long, flesh coloured, with three coloured
floral leaves.
Empalement. Cup four-leaved, which are of a broad oval form, flesh
coloured, and longer than the blossom.
The Blossom is small, of a pitcher-shape, light flesh colour at the end,
greenish at the base; the segments of the mouth are bent back, and deep
flesh coloured.
Chives. Eight hair-like threads; the tips crested, and within the blossom.
Pointal. Seed-vessel nearly globular; the shaft thread-shaped, partly out
of the blossom. Summit four-cornered.
Native of the Cape of Good Hope.
Flowers from September, till December.
REFERENCE.
1. The Empalement, and Blossom.

2. The Blossom.
3. The Empalement magnified.
4. The Chives and Pointal.
5. The Chives detached from the Pointal; one tip magnified.
6. The Shaft and its Summit magnified.
ERICA campanulata.
CHARACTER SPECIFICUS.
Erica, antheris muticis, inclusis; floribus solitariis, cernuis; calycibus

coloratis; corollis globoso-campanulatis, luteis; foliis quaternis, linearibus,
angustissimis.
DESCRIPTIO.
Caulis fruticosus, pedalis, erectus, filiformis; rami et ramuli filiformes,

virgati, glabri.
Folia quaterna, linearia, angusta, glabra, erecta; petiolis brevissimis,
adpressis.
Flores terminales, subsolitarii, cernui; pedunculi colorati, bracteis tribus,
coloratis, subulatis, instructi.
Calyx. Perianthium tetraphyllum, foliolis ovatis, acuminatis, concavis,
adpressis, coloratis.
Corolla globoso-campanulata, lutea; laciniis semi-ovatis, subreflexis.
Stamina. Filamenta octo, capillaria, incurvata. Antheræ muticæ, inclusæ.
Pistillum. Germen sub-globosum. Stylus columnaris. Stigma
tetragonum.
Habitat ad Caput Bonæ Spei.
Floret a mensi Junii, in Augustum.
REFERENTIA.
1. Calyx, et Corolla.
2. Corolla.
3. Calyx, lente auctus.
4. Stamina, et Pistillum.
5. Anthera una, aucta.
6. Pistillum, lente auctum.
SPECIFIC CHARACTER.
Heath, with beardless tips, within the blossom; flowers grow singly,
hanging down; cups coloured; blossoms globularly bell-shaped, and yellow;
leaves grow by fours, are linear, and very narrow.
DESCRIPTION.
Stem shrubby, grows a foot high, upright, and thread-shaped; the greater
and smaller branches are thread-shaped, twiggy, and smooth.
Leaves grow by fours, linear, narrow, smooth, and upright; very short
foot-stalks, pressed to the branches.
Flowers grow mostly solitary, at the end of the smaller branches,
hanging down; the foot-stalks are coloured, having three awl-shaped,
coloured floral leaves on them.
Empalement. Cup four-leaved, leaflets egg-shaped, pointed, concave,
pressed to the blossom, and coloured.
Blossom, globularly bell-shaped, and yellow; the segments half egg-
shaped, and a little reflexed.
Chives. Eight hair-like threads, curved inwards. Tips beardless, and
within the blossom.
Pointal. Seed-bud nearly globe-shaped. Shaft pillar-shaped. Summit
four-cornered.
Flowers from June, till August.
REFERENCE.

2. The Blossom.
3. The Empalement, magnified.
4. The Chives, and Pointal.
5. A Tip, magnified.
6. The Pointal, magnified.
ERICA capitata.
Erica, antheris muticis, sub-inclusis; corollis globosis, albidis, tomentosis,

tectis calyce magno, hispido, colorato; foliis ternis, pilosis.
DESCRIPTIO.
Caulis erectus, laxus, filiformis, fuseus; rami filiformes, villosi; ramuli

capillares, frequentes, tomentosi.
Folia terna, linearia, obtusa, dorso sulcata, pilis longis hirta; petiolis
brevissimis, adpressis.
Flores in extremis ramulis terminales bini, vel terni, cernuo-patenti;
pedunculi brevissimi, bracteis tribus, minutis, adpressis, instructi.
Calyx. Perianthium tetraphyllum, foliolis ovatis, concavis, incurvatis
maximis, totis densissime hirtis, adpressis, luteo-viridis.
Corolla sub-globosa, alba, lanata, in sinu calycis fere recondita.
Stamina. Filamenta octo capillaria, receptaculo inserta. Antheræ muticæ
sub-inclusæ.
Pistillum. Germen subrotundum, apice pilosum. Stylus filiformis,
exsertus. Stigma tetragonum.
Floret a mensi Julii, in Octobrem.
REFERENTIA.
2. Corolla.
5. Stamina a Pistillo diducta, anthera una lente aucta.
6. Stylus, et Stigma, lente aucta.
SPECIFIC CHARACTER.
Heath, with beardless tips, just within the blossoms, which are globular,
white and downy, being covered with a large, hairy, coloured cup; leaves
grow by threes, and are hairy.
DESCRIPTION.
Stem upright, weak, thread-shaped, and brown; branches thread-shaped,

and hairy; small branches like hairs, numerous, and downy.
Leaves grow by threes, linear, blunt, furrowed at the back, and covered
with long, harsh hairs; foot-stalks very short, and pressed to the branches.
Flowers grow at the extremity of the smaller branches, by twos or
threes, spreading out, and nodding; foot-stalks very short, having three
small floral leaves, which are pressed to the blossom.
Empalement. Cup of four leaves, which are egg-shaped, concave, turned
inwards, very large, quite covered with strong hairs, pressed to the
blossoms, and of a yellow-green.
Blossom nearly globular, white and woolly, almost hid within the cup.
Chives. Eight hair-like threads, fixed into the receptacle. Tips beardless,
and nearly within the blossom.
Pointal. Seed-bud nearly round, and hairy at the end. Shaft thread-
shaped, and without the blossom. Summit four-cornered.
Flowers from the month of July, till October.
REFERENCE.
1. Empalement, and Blossom.

2. A Blossom.
5. The Chives detached from the Pointal, one Tip magnified.
6. The Shaft, and its Summit, magnified.
ERICA cerinthoides.
Erica, antheris muticis, inclusis; corollis tubulato-ventricosis, læte

sanguineis, fasciculatis, hispidis; foliis quaternis, rigidis, ciliatis.
DESCRIPTIO.
Caulis flexibilis, erectus, cinereus, sesquipedalis; rami pauci, erecti, raro

ramulosi.
Folia quaterna, ciliata, oblonga, convexa, subtus sulco exerata, petiolis
brevibus, adpressis.
Flores magni, sessiles, aggesti in capitulum, cernui, pedunculi hispidi,
bracteis tribus foliis similibus instructi.
Calyx. Perianthium tetraphyllum, foliolis lanceolatis, hispidis, foliis
similibus.
Corolla, tubulato-ventricosa, læte sanguinea, hirsuta, ore obsolete
quadrifida; pollicaria.
Stamina. Filamenta octo capillaria. Antheræ muticæ, inclusæ.
Pistillum. Germen cylindricum, hirsutum. Stylus filiformis, sub-
exsertus. Stigma tetragonum.
Floret a mensi Augusti, in Aprilem.
REFERENTIA.
2. Calyx lente auctus.
4. Stamina a Pistillo diducta; anthera una lente aucta.
5. Stylus, et Stigma, lente aucta.
SPECIFIC CHARACTER.
Heath, with beardless tips, within the blossoms, which are of an inflated
tubular shape, of a rich blood colour, hairy, and bundled together; the leaves
grow by fours, are harsh, and lashed.
DESCRIPTION.
Stem grows upright, pliant, ash-coloured, and a foot and a half high; the
branches are few, and upright, seldom branching.
The Leaves grow by fours, are lashed, oblong, rounded on the upper
surface, and deeply furrowed on the under side, with short foot-stalks,
pressed to the stems.
The Flowers are large, growing in close bunches, fixed altogether at the
end of the branches, bending downward; the foot-stalks are hairy, having
three floral leaves, similar to the other leaves.
Empalement. Cup four-leaved, which are lance-shaped, hairy, and like
the other leaves.
Blossom, of an inflated tubular form, hairy, and of a rich red or blood
colour, the mouth slightly cut into four segments; an inch long.
Chives. Eight hair-like threads. Tips beardless, and within the blossom.
Pointal. Seed-vessel cylinder shape, and hairy. Style thread-shaped,
nearly without the blossom. Summit four-cornered.
Flowers from August, till April.
REFERENCE.

2. The Empalement magnified.
5. The Shaft, and its Summit, magnified.
ERICA cernua.
Erica, antheris cristatis, inclusis; floribus umbellatis, cernuis, secundis, sub-

ovatis, pallide-carneis; foliis quaternis.
DESCRIPTIO.
Caulis fruticosus, erectus, pedalis; rami sub-simplices, erecto-patenti.

Folia quaterna, linearia, obtusa, subtus sulcata, sub-scabrida; petiolis
brevissimis, adpressis.
Flores in apice ramorem umbellati, cernui; pedunculi longi, colorati,
bracteis tribus, linearibus, instructi.
Calyx. Perianthium tetraphyllum, foliolis minutis, subulatis, ciliatis,
coloratis.
Corolla sub-ovata, pallide-carnea; laciniis limbi acuminatis, sub-
erectis.
Stamina. Filamenta octo capillaria apice introrsum declinata. Antheræ
cristatæ, inclusæ.
Pistillum. Germen turbinatum, fulcatum. Stylus cylindricus, sub-
inclusus. Stigma obsolete tetragonum.
Floret a mensi Augusti in Decembrem.
REFERENTIA.
3. Stamina a Pistillo diducta, anthera una lente aucta.
4. Germen, Stylus, et Stigma, lente aucta.
SPECIFIC CHARACTER.
Heath, with crested tips, within the blossom; the flowers grow in bunches,
nodding, all pointing one way, nearly egg-shaped, and of a pale flesh colour;
leaves growing by fours.
DESCRIPTION.
Stem shrubby, upright, and grows a foot high; the branches are almost
simple, upright, and spreading.
Leaves grow by fours, linear, blunt, channelled beneath, and roughish;
having very short foot-stalks pressed to the branches.
Flowers grow in bunches, at the end of the branches, nodding; the foot-
stalks are long and coloured, with three linear floral-leaves on them.
Empalement. Cup of four leaves, which are small, awl-shaped, fringed,
and coloured.
Blossom, nearly egg-shaped, of a pale flesh-colour; the segments of the
border tapered, and nearly upright.
Chives. Eight hair-like threads, bent downward on the inner side. Tips
crested, and within the blossom.
Pointal. Seed-bud turban-shaped, and channelled. Shaft cylindrical just
within the blossom. Summit obscurely four-cornered.
Flowers from August, till December.
REFERENCE.

4. The Seed-bud, Shaft, and Summit, magnified.
A SHORT DISSERTATION, &c.
Antecedent to the year 1772, the few species of this, now so numerous
Genus, known in our British gardens, were, the E. vulgaris, E. Tetralix, E.
cineria and E. vagans natives; the E. Dabœcii, from Ireland; the E. arborea
introduced in 1748, from Madeira; the E. herbacea or carnea in 1763, from
Switzerland; the E. mediterranea in 1765, from Minorca, and the E.
scoparia, E. viridi-purpurea, E. australis, E. ciliaris and E. umbellata, from
Portugal, between the years 1768 and 1707. The two other European species
we possess, the E. stricta and E. multiflora, natives of Spain, have been but
twelve years in cultivation with us. Of the African species, found within the
district of the Cape of Good Hope and the adjacent territory, which have
swelled the Genus to so great an extent; and which, but as an echo to the
general voice, may be said to contribute, by the extreme brilliancy of the
flowers of these species, more than any other, to the present splendor of our
green-houses, were unknown, till the above æra, to our English botanists,
but by name. In the year 1771 seeds of two species were received, at the
Hammersmith nursery, from the Cape, both of them vegetated; the first
which flowered, proving the E. tubiflora, of the Sp. Plant. of Linnæus; the
other, from the resemblance it bears to the Spruce Fir, was then named E.
abietina; but since, altered in the Kew catalogue, to E. concinna. Two years
subsequent, 1774, Mr. Francis Masson, botanical collector to His Majesty at
the Cape, laid the foundation for the celebrity of that superb collection at
Kew, which for many years, with unrivalled lustre, far outshone all others,
particularly by the number and variety of this most beautiful tribe of plants:
for which we refer to the second Vol. of the catalogue of that garden; where
the E. curviflora, E. lutea, E. cruenta, E. persoluta, E. baccans, E. marifolia,
E. abietina, E. corifolia, E. paniculata, E. empetrifolia, E. spumosa, E.
capitata, E. conspicua, E. cerinthoides, E. viscaria, E. Plukenetii, E. Petiveri,
and E. petiolata, are all stated to have been of that year’s introduction. From
this period, till within these few years, the accession was so rapid, that it
would be difficult, nay nearly impracticable, to ascertain the precise date
when most of the remaining species were introduced; as many different
collectors were about this time, or shortly after, producing in their
collections new species to which they each claimed the honour of priority of
introduction; the enumeration of these, therefore, in succession would be but

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Molybdenum and Tungsten Enzymes

RSC Metallobiology Series

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Molybdenum and Tungsten

Spectroscopic and Theoretical Investigations

RSC Metallobiology Series No. 7

Print ISBN: 978-1-78262-878-1

© The Royal Society of Chemistry 2017

All rights reserved

Published by The Royal Society of Chemistry,

Registered Charity Number 207890

For further information see our web site at www.rsc.org

RSC Metallobiology Series No. 7

a wide range of metabolic pathways and play particularly prominent roles

RSC Metallobiology Series No. 7

and molybdenum complexes when, beginning in the late 1970s, he became

Chapter 1 Spectroscopic and Electronic Structure Studies

Chapter 2 Spectroscopic and Electronic Structure Studies of

2.1 I ntroduction and Scope  13

RSC Metallobiology Series No. 7

2.4 Xanthine Oxidoreductase (XOR)  35

Chapter 3 Electron Paramagnetic Resonance Studies of

3.3.3 g-Tensor Analysis of Mo-bisPGD Active Site:

Chapter 4 X-Ray Absorption Spectroscopy of Molybdenum and

4.1 I ntroduction  121

4.3.4 Combining XAS with Other

Chapter 5 Electrochemistry of Molybdenum and Tungsten

5.1 I ntroduction  168

6.1 I ntroduction  223

Chapter 7 Computational Studies of Molybdenum and Tungsten

7.1 I ntroduction  275

7.7 Conclusions  314

Subject Index  322

Spectroscopic and Electronic

RSC Metallobiology Series No. 7

detail how spectroscopy, structure, electrochemistry and theory have been

Spectroscopic and Electronic Structure Studies Probing Mechanism 3

E-MoVI + R + H2O → E-MoIV + R–O + 2H+

Regarding substrate oxidations, reductive oxygen atom transfer is given

E-MoVI + R–H + H2O → E-MoIV + R–OH + 2H+

Spectroscopic and Electronic Structure Studies Probing Mechanism 5

crucial interrelationships between spectroscopic features of the active sites

Spectroscopic and Electronic Structure Studies Probing Mechanism 7

canonical members of each pyranopterin Mo enzyme family, as well as the

tein electrochemistry of pyranopterin Mo enzymes that begins with a tutorial

N2 + 8e− + 16MgATP + 8H+ → 2NH3 + H2 + 16MgADP + 16Pi

The catalytic mechanism of nitrogenase is discussed in terms of the Lowe–

Spectroscopic and Electronic Structure Studies Probing Mechanism 9

Figure 1.5  modified Lowe–Thorneley scheme for nitrogenase. Adapted from

exchange coupled paramagnetic spin centers. For nitrogenase, Mössbauer

numerous contributions from researchers engaged in the biological, struc-

Chapter 1 Spectroscopic and Electronic Structure Studies

Chapter 2 Spectroscopic and Electronic Structure Studies of

2.1 I ntroduction and Scope 13

2.4 Xanthine Oxidoreductase (XOR) 35

Chapter 3 Electron Paramagnetic Resonance Studies of

3.3.3 g-Tensor Analysis of Mo-bisPGD Active Site:

Chapter 4 X-Ray Absorption Spectroscopy of Molybdenum and

4.1 I ntroduction 121

4.3.4 Combining XAS with Other

Chapter 5 Electrochemistry of Molybdenum and Tungsten

5.1 I ntroduction 168

6.1 I ntroduction 223

Chapter 7 Computational Studies of Molybdenum and Tungsten

7.1 I ntroduction 275

7.7 Conclusions 314

Subject Index 322

Figure 1.5 modified Lowe–Thorneley scheme for nitrogenase. Adapted from

22. G. J. Workun, K. Moquin, R. A. Rothery and J. H. Weiner, Microbiol. Mol.

2.1 Introduction and Scope