Profiling the Active Site of a Copper Enzyme through Its Far‐Infrared Fingerprint

Marboutin, Laure; Petitjean, Hugo; Xerri, Bertrand; Vita, Nicolas; Dupeyrat, François; Flament, Jean‐Pierre; Berthomieu, Dorothée; Berthomieu, Catherine

doi:10.1002/anie.201102014

Cited by 8 publications

(13 citation statements)

References 47 publications

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“…However, no contributions of the histidine side-chain modes are predicted in this domain. These bands may arise from contributions involving peptide bonds or from modes involving metal-histi- [8] The experimental shifts observed upon 15 N labelling support these assignments as they are close to those predicted for these modes (Table S5).…”

Section: Experimental Ftir Difference Spectra Of Cuzn-sodsupporting

confidence: 79%

“…This modelling approach was demonstrated as robust and accurate for the prediction and assignment of experimental low-frequency IR modes of Cu-histamine complexes. [8,15] Self-consistent field (SCF) cycles and geometry optimizations were considered as converged when energy changes were less than 10 -8 and 10…”

Section: Methodsmentioning

confidence: 99%

“…IR data have been obtained for the Cu II and Cu I redox states in the mid-IR to far-IR range. [8,14] Figure 1. Cu II ,Zn-SOD and Cu I ,Zn-SOD metal site cluster structures used as 8H and 9H (see the "Theoretical Calculations" section).…”

Section: Introductionmentioning

confidence: 97%

“…The recent development of light‐induced and electrochemically induced FTIR difference spectroscopy in the far‐infrared domain (in which metal–ligand bonds vibrate) opens the way to analyze a large number of metal sites by infrared spectroscopy. These data concern the manganese complex of photosystem II (PS II),1–7 the copper centres of superoxide dismutase8 or azurin,9 the heme iron of cytochrome and related models10,11 and iron–sulfur centres 12. The analysis of these data in terms of ligand and metal–ligand bond properties necessitates a comparison with those of well‐characterized model compounds as well as detailed analysis of the normal modes by theoretical approaches.…”

Section: Introductionmentioning

confidence: 99%

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Mid‐ and Far‐Infrared Marker Bands of the Metal Coordination Sites of the Histidine Side Chains in the Protein Cu,Zn‐Superoxide Dismutase

et al. 2014

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International audienceVibrational spectroscopy gives important information on the properties of ligand and metal–ligand bonds in metalloenzymes. Infrared spectroscopy is appealing for the study of metal active sites that are not amenable to Raman spectroscopy. We present a combined experimental and theoretical approach to analyze the mid- and far-IR spectra of Cu,Zn-superoxide dismutase (Cu,Zn-SOD) as a probe of the histidine ligands. This metalloenzyme provides a unique model to identify specific IR signatures of metal–histidine coordination and to study their alterations as a function of the metal (copper/zinc), the copper valence state (+I/+II), the histidine coordination mode (Nτ and Nπ) and the histidine protonation state. DFT calculations combined with normal mode descriptions from potential energy distribution calculations were performed on two slightly different cluster models. Differences in the constraints at the side chain of one histidine Cu ligand sensibly modify the geometric parameters and vibrational properties. Electrochemically induced FTIR difference spectroscopy provided mid- and far-IR fingerprint spectra of the Cu protein in aqueous media that are sensitive to the redox state of the Cu centre at the active site. Comparisons of the DFT predictions with the experimental IR modes of the histidine ligands at the Cu,Zn-SOD active site showed that useful mid-IR markers of histidine Nτ and Nπ coordination were predicted with good accuracy. The DFT analysis further demonstrated a link between the ν(C4–C5) mode frequency of His46 and the specific properties of the His46–Cu bond in Cu,Zn-SOD. A combined theoretical and experimental approach on samples in H2O and 2H2O or 15N-labelled samples identified the contributions from the histidine side chain modes in the 669–629 cm–1 region

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Section: Experimental Ftir Difference Spectra Of Cuzn-sodsupporting

confidence: 79%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

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Mid‐ and Far‐Infrared Marker Bands of the Metal Coordination Sites of the Histidine Side Chains in the Protein Cu,Zn‐Superoxide Dismutase

et al. 2014

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“…The concept of the transmission cell is not significantly changed for the mid-infrared (MIR) spectral range. In the far infrared (below 600 cm -1 ), where metal-ligand vibrations can be observed, a cell with silicon or diamond windows was created on the basis of the same cell geometry (El Khoury and Hellwig , 2011 ;Marboutin et al , 2011 ). This technique is complementary to metal-ligand studies performed by coupled electrochemical approaches and Raman spectroscopies in this spectral range.…”

Section: Type Of Cellsmentioning

confidence: 99%

Recent advances in the electrochemistry and spectroelectrochemistry of membrane proteins

Melin

Hellwig

2013

Biological Chemistry

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Abstract:Integral membrane proteins are encountered in fundamental natural processes, such as photosynthesis and respiration. The relation between the structure of the proteins and their function and dynamics are still not clear in most cases. Once fully understood, these processes could ultimately help researchers to develop alternative methods for producing energy, either from light or biomass. They could also lead to more efficient antibiotics, which would selectively inhibit a specific membrane protein of pathogenic bacteria. Since the chemical reactions involved in both photosynthesis and respiration are redox reactions, electrochemical methods can play a considerable role in uncovering their mechanisms. The electrochemical characterization of membrane proteins is, however, quite challenging. An overview on the techniques used for the characterization of membrane proteins, including classical approaches such as voltammetry and spectroelectrochemistry, and recent developments, such as their combination with surface-enhanced techniques is given.

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In Situ and Operando Techniques for Investigating Electron Transfer in Biological Systems

et al. 2020

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When bioelectrochemistry meets other analytical techniques for in situ experiments, new possibilities for understanding the nature of charge‐transfer reactions are provided. Despite progress in recent years in analytical instrumentation, a number of key issues remains for bioelectrochemistry, in terms of the mechanistic description of the surface reactions involved in biomolecules’ electron transfer. The main challenges of these techniques are the concomitant detection of electron transfer and molecular alteration as well as the development of suitable bioelectrodes. This review discusses the most recent and emerging in situ and operando electrochemical methods used to study biological systems such as redox proteins, nucleic acids, small biomolecules, cells, and biomembranes, focusing on electrochemical methods coupled with spectroscopic, spectrometric, and microscopic techniques.

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Profiling the Active Site of a Copper Enzyme through Its Far‐Infrared Fingerprint

Cited by 8 publications

References 47 publications

Mid‐ and Far‐Infrared Marker Bands of the Metal Coordination Sites of the Histidine Side Chains in the Protein Cu,Zn‐Superoxide Dismutase

Mid‐ and Far‐Infrared Marker Bands of the Metal Coordination Sites of the Histidine Side Chains in the Protein Cu,Zn‐Superoxide Dismutase

Recent advances in the electrochemistry and spectroelectrochemistry of membrane proteins

In Situ and Operando Techniques for Investigating Electron Transfer in Biological Systems

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