A large-scale comparative analysis of affinity, thermodynamics and functional characteristics of twelve cytochrome P450 isoforms and their redox partners.

A large-scale comparative analysis of affinity, thermodynamics and functional characteristics of twelve cytochrome P450 isoforms and their redox partners.
Biochimie. 2019 Apr 26;:
Authors: Yablokov ЕО, Sushko ТА, Ershov PV, Florinskaya АV, Gnedenko ОV, Shkel ТV, Grabovec IP, Strushkevich NV, Kaluzhskiy LA, Usanov SА, Gilep АА, Ivanov АS
Abstract
The aim of the present work was to establish the thermodynamic and functional differences in the protein-protein interactions between the components of the P450-dependent mitochondrial (mit) and microsomal (mic) monooxygenase systems using 12 different isoforms of cytochromes P450 and two redox partners, NADPH-dependent cytochrome P450 reductase (CPR) and adrenodoxin (Adx). Comparative analysis of the affinity, thermodynamics, enzymatic activity and the ability for one-electron reduction has been carried out. The study of protein-protein interactions to determine the equilibrium dissociation constants (Kd) was performed using surface plasmon resonance (SPR) biosensor Biacore 3000. We demonstrated that CPR and Adx interacted with both, micCYPs and mitCYPs, with different affinities (Kd values ranged from 0.01 to 2 μM). All complexes of microsomal (micCYP) and mitochondrial (mitCYP) cytochrome P450 with redox partners can be divided into three groups depending on the prevalent role of either enthalpy or entropy contribution. About 90% of CYP/redox partner complexes were entropy-driv...

Source: Biochimie - April 25, 2019 Category: Biochemistry Authors: Yablokov ЕО, Sushko ТА, Ershov PV, Florinskaya АV, Gnedenko ОV, Shkel ТV, Grabovec IP, Strushkevich NV, Kaluzhskiy LA, Usanov SА, Gilep АА, Ivanov АS Tags: Biochimie Source Type: research