Direct electron transfer between cytochrome P450scc and gold nanoparticles on screen-printed rhodium–graphite electrodes

Shumyantseva, Victoria V.; Carrara, Sandro; Bavastrello, Valter; Riley, D. Jason; Булко, Т. В.; Skryabin, K. G.; Archakov, Alexander I.; Nicolini, Claudio

doi:10.1016/j.bios.2004.10.008

Cited by 109 publications

(71 citation statements)

References 27 publications

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“…Complicating these systems, the active redox site is often deeply embedded in the interior of the enzyme, insulating it from the electrode and impeding electron transfer between the electrode and the redox site of enzyme. Finally, immobilization by direct attachment may impede or prohibit substrate binding or metabolite release through steric constraints (Habermüller et al, 2000;Joseph et al, 2003;Shumyantseva et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Electrocatalytic Drug Metabolism by CYP2C9 Bonded to A Self-Assembled Monolayer-Modified Electrode

Yang¹,

Kabulski²,

Wollenberg³

et al. 2009

Drug Metab Dispos

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ABSTRACT:Cytochrome P450 (P450) enzymes typically require the presence of at least cytochrome P450 reductase (CPR) and NADPH to carry out the metabolism of xenobiotics. To address whether the need for redox transfer proteins and the NADPH cofactor protein could be obviated, CYP2C9 was bonded to a gold electrode through an 11-mercaptoundecanoic acid and octanethiol self-assembled monolayer (SAM) through which a current could be applied. Cyclic voltammetry demonstrated direct electrochemistry of the CYP2C9 enzyme bonded to the electrode and fast electron transfer between the heme iron and the gold electrode. To determine whether this system could metabolize warfarin analogous to microsomal or expressed enzyme systems containing CYP2C9, warfarin was incubated with the CYP2C9-SAM-gold electrode and a controlled potential was applied.The expected 7-hydroxywarfarin metabolite was observed, analogous to expressed CYP2C9 systems, wherein this is the predominant metabolite. Current-concentration data generated with increasing concentrations of warfarin were used to determine the MichaelisMenten constant (K m ) for the hydroxylation of warfarin (3 M), which is in good agreement with previous literature regarding K m values for this reaction. In summary, the CYP2C9-SAM-gold electrode system was able to carry out the metabolism of warfarin only after application of an electrical potential, but in the absence of either CPR or NADPH. Furthermore, this system may provide a unique platform for both studying P450 enzyme electrochemistry and as a bioreactor to produce metabolites without the need for expensive redox transfer proteins and cofactors.Cytochrome P450 (P450) enzymes are important for drug metabolism in humans, accounting for ϳ75% metabolism of drugs (Williams et al., 2004). Numerous factors affect the P450 metabolism rate and the resulting metabolite structure including electron transfer, protein-protein interactions, concentration and structure of the substrate, and the source and specific means whereby the P450 was prepared (Guengerich, 2007). In practice, it is difficult to isolate the individual contributions of these factors because they are often interdependent. Thus, it is of interest to devise model platforms that can be used to independently control these parameters to monitor their respective effects on metabolism.P450 catalysis requires a constant supply of NADPH as the electron source and cytochrome P450 reductase (CPR) to deliver the electrons.In attempts to obviate the requirement of these redox partners/cofactors for catalysis, P450 enzymes have been immobilized on electrodes so that the electrode supplies electrons to drive the P450 catalytic cycle (Estabrook et al., 1996;Reipa et al., 1997;Gilardi et al., 2002) with effective electrical communication between the electrode and the enzyme being critical (Yang et al., 2008). Direct adsorption of enzymes on bare electrodes, such as gold, platinum, and graphite, results in diminished biocatalytic activity (Habermüller et al., 2000;Joseph et al., 2003;Shumy...

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Section: Introductionmentioning

confidence: 99%

Electrocatalytic Drug Metabolism by CYP2C9 Bonded to A Self-Assembled Monolayer-Modified Electrode

Yang¹,

Kabulski²,

Wollenberg³

et al. 2009

Drug Metab Dispos

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“…The immobilization of protein on gold nanoparticles can help the protein to keep a favoured orientation or to create conducting channels between the prosthetic groups and the electrode surface, thus reducing the effective electron transfer distance (Wu & Hu, 2007). It has been demonstrated (Shumyantseva, Carrara, Bavastrello, Riley, Bulko, Skryabin, Archakov, Nicolini, 2005) that gold nanoparticles improve the sensitivity of cholesterol biosensor with cytochrome P450scc. Other studies Peng et al, 2008) showed the feasibility in enhance redox current of CYP2B6 to be incorporated into films made by zirconium dioxide nanoparticles and platinum components and into a chitosan modified colloidal gold nanoparticles .…”

Section: Cyps Immobilization Techniquesmentioning

confidence: 99%

P450-Based Nano-Bio-Sensors for Personalized Medicine

Baj-Rossi¹,

Micheli²,

Carrara³

2011

Biosensors - Emerging Materials and Applications

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“…With the advent of nanotechnology, an interest was focused on developing new nanomaterials and to employ them for an improved sensor sensitivity and for a preservation of the native structure of the enzyme when immobilized onto the electrode surface [18][19][20][21][22][23][24][25]. In particular, more recent studies demonstrate that the electrode modification with metal nanoparticles such as gold [4,13,14] or platinum [15][16][17] decreases the overpotential for the detection of H 2 O 2 . Nanostructures based on transition metals such as Au and Pt, thanks to their capability to enhance the electrocatalytic activity towards H 2 O 2 oxidation, have been incorporated into electrode surfaces to make biosensors [26][27][28][29][30].…”

Section: Introductionmentioning

confidence: 99%

A bimetallic nanocoral Au decorated with Pt nanoflowers (bio)sensor for H2O2 detection at low potential

Sanzò

Taurino

Puppo

et al. 2017

Methods

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a b s t r a c tIn this work, we have developed for the first time a method to make novel gold and platinum hybrid bimetallic nanostructures differing in shape and size. Au-Pt nanostructures were prepared by electrodeposition in two simple steps. The first step consists of the electrodeposition of nanocoral Au onto a gold substrate using hydrogen as a dynamic template in an ammonium chloride solution. After that, the Pt nanostructures were deposited onto the nanocoral Au organized in pores. Using Pt (II) and Pt (IV), we realized nanocoral Au decorated with Pt nanospheres and nanocoral Au decorated with Pt nanoflowers, respectively. The bimetallic nanostructures showed better capability to electrochemically oxidize hydrogen peroxide compared with nanocoral Au. Moreover, Au-Pt nanostructures were able to lower the potential of detection and a higher performance was obtained at a low applied potential. Then, glucose oxidase was immobilized onto the bimetallic Au-Pt nanostructure using cross-linking with glutaraldehyde. The biosensor was characterized by chronoamperometry at +0.15 V vs. Ag pseudo-reference electrode (PRE) and showed good analytical performances with a linear range from 0.01 to 2.00 mM and a sensitivity of 33.66 mA/mM cm 2 . The good value of K m app (2.28 mM) demonstrates that the hybrid nanostructure is a favorable environment for the enzyme. Moreover, the low working potential can minimize the interference from ascorbic acid and uric acid as well as reducing power consumption to effect sensing. The simple procedure to realize this nanostructure and to immobilize enzymes, as well as the analytical performances of the resulting devices, encourage the use of this technology for the development of biosensors for clinical analysis.

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Direct electron transfer between cytochrome P450scc and gold nanoparticles on screen-printed rhodium–graphite electrodes

Cited by 109 publications

References 27 publications

Electrocatalytic Drug Metabolism by CYP2C9 Bonded to A Self-Assembled Monolayer-Modified Electrode

Electrocatalytic Drug Metabolism by CYP2C9 Bonded to A Self-Assembled Monolayer-Modified Electrode

P450-Based Nano-Bio-Sensors for Personalized Medicine

A bimetallic nanocoral Au decorated with Pt nanoflowers (bio)sensor for H2O2 detection at low potential

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