TY - JOUR
T1 - Electrochemical Activity of Cytochrome P450 1A2
T2 - The Relevance of O2 Control and the Natural Electron Donor
AU - Silveira, Célia M.
AU - Rodrigues, Patrícia R.
AU - Ghach, Wissam
AU - Pereira, Sofia A.
AU - Esteves, Francisco
AU - Kranendonk, Michel
AU - Etienne, Mathieu
AU - Almeida, M. Gabriela
N1 - info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F04585%2F2020/PT#
i3N UID/CTM/50025/2013)#
info:eu-repo/grantAgreement/FCT/OE/SFRH%2FBPD%2F79566%2F2011/PT#
info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UID%2FBIM%2F00009%2F2019/PT#
co‐financed by the ERDF under the PT2020 Partnership Agreement (POCI‐01‐0145‐FEDER‐007728). and from Project LISBOA‐01‐0145‐ FEDER‐007660 (Microbiologia Molecular, Estrutural e Celular), which is funded by FEDER funds through COMPETE 2020‐Programa Operacional Competitividade e Internacionalização (POCI).
PY - 2021/2/1
Y1 - 2021/2/1
N2 - The direct electrochemical response of membrane-bound human cytochrome P450 1A2 (CYP1A2) was studied on pyrolytic graphite electrodes, while encapsulated in a sol-gel matrix. The enzymatic reduction of O2 was evaluated in the presence and absence of its electron donor partner, cytochrome P450 oxidoreductase (CPR). When used without CPR, CYP1A2 was shown to be highly sensitive to O2 even in the presence of residual amounts. Under aerobic conditions (air-saturated solutions), the catalytic signal attributed to the reaction with O2 was lost, suggesting the enzyme was inactivated. In contrast, the CYP1A2/CPR complex retained O2 reductase activity with high O2 concentration in solution. The results demonstrated a crucial role of CPR in stabilizing the immobilized CYP1A2 enzyme and in the preservation of O2 electrocatalysis, when using this electrochemical set-up. Though the enzyme's monooxygenase activity towards caffeine was not detected, this study highlights the complexity of coupling CYP1A2 reduction currents with substrate turnover, owing to the simultaneous electrochemical measurement of the O2 reduction reaction.
AB - The direct electrochemical response of membrane-bound human cytochrome P450 1A2 (CYP1A2) was studied on pyrolytic graphite electrodes, while encapsulated in a sol-gel matrix. The enzymatic reduction of O2 was evaluated in the presence and absence of its electron donor partner, cytochrome P450 oxidoreductase (CPR). When used without CPR, CYP1A2 was shown to be highly sensitive to O2 even in the presence of residual amounts. Under aerobic conditions (air-saturated solutions), the catalytic signal attributed to the reaction with O2 was lost, suggesting the enzyme was inactivated. In contrast, the CYP1A2/CPR complex retained O2 reductase activity with high O2 concentration in solution. The results demonstrated a crucial role of CPR in stabilizing the immobilized CYP1A2 enzyme and in the preservation of O2 electrocatalysis, when using this electrochemical set-up. Though the enzyme's monooxygenase activity towards caffeine was not detected, this study highlights the complexity of coupling CYP1A2 reduction currents with substrate turnover, owing to the simultaneous electrochemical measurement of the O2 reduction reaction.
KW - CYP electrochemistry
KW - cytochrome P450
KW - cytochrome P450 oxidoreductase
KW - O reduction
KW - sol-gel
UR - http://www.scopus.com/inward/record.url?scp=85097796934&partnerID=8YFLogxK
U2 - 10.1002/celc.202001255
DO - 10.1002/celc.202001255
M3 - Article
AN - SCOPUS:85097796934
SN - 2196-0216
VL - 8
SP - 500
EP - 507
JO - Chemelectrochem
JF - Chemelectrochem
IS - 3
ER -