Engineered human CYP2C9 and its main polymorphic variants for bioelectrochemical measurements of catalytic response
| Autor: | Gianfranco Gilardi, Silvia Castrignanò, Sheila J. Sadeghi, Paola Panicco, Giovanna Di Nardo |
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| Rok vydání: | 2020 |
| Předmět: |
Flavodoxin
Biophysics 02 engineering and technology Glassy carbon Protein Engineering 01 natural sciences Redox Cytochrome P450 2C9 Personalized medicine Polymorphism Warfarin Electrochemistry Humans Physical and Theoretical Chemistry Cytochrome P-450 CYP2C9 chemistry.chemical_classification Anticoagulant drug biology fungi 010401 analytical chemistry Wild type General Medicine 021001 nanoscience & nanotechnology Enzymes Immobilized 0104 chemical sciences Enzyme chemistry Biochemistry biology.protein Biocatalysis Cyclic voltammetry 0210 nano-technology Drug metabolism |
| Zdroj: | Bioelectrochemistry (Amsterdam, Netherlands). 138 |
| ISSN: | 1878-562X |
| Popis: | Polymorphism is an important aspect in drug metabolism responsible for different individual response to drug dosage, often leading to adverse drug reactions. Here human CYP2C9 as well as its polymorphic variants CYP2C9*2 and CYP2C9*3 present in approximately 35% of the Caucasian population have been engineered by linking their gene to the one of D. vulgaris flavodoxin (FLD) that acts as regulator of the electron flow from the electrode surface to the haem. The redox properties of the immobilised proteins were investigated by cyclic voltammetry and electrocatalysis was measured in presence of the largely used anticoagulant drug S-warfarin, marker substrate for CYP2C9. Immobilisation of the CYP2C9-FLD, CYP2C9*2-FLD and CYP2C9*3-FLD on DDAB modified glassy carbon electrodes showed well defined redox couples on the oxygen-free cyclic voltammograms and mid-point potentials of all enzymes were calculated. Electrocatalysis in presence of substrate and quantification of the product formed showed lower catalytic activities for the CYP2C9*3-FLD (2.73 ± 1.07 min−1) and CYP2C9*2-FLD (12.42 ± 2.17 min−1) compared to the wild type CYP2C9-FLD (18.23 ± 1.29 min−1). These differences in activity among the CYP2C9 variants are in line with the reported literature data, and this set the basis for the use of the bio-electrode for the measurement of the different catalytic responses towards drugs very relevant in therapy. |
| Databáze: | OpenAIRE |
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