Aluminum-substituted heme domain of P450BM-3 (BMP): Introducing a heme-derived fluorescent probe for studies of substrate binding and protein-protein interactions in cytochromes P450

2013 ◽  
Vol 60 (1) ◽  
pp. 41-51 ◽  
Author(s):  
Dmitri R. Davydov ◽  
Gelii V. Ponomarev ◽  
Ekaterina Bobrovnikova-Marjon ◽  
Donovan C. Haines ◽  
Julian A. Peterson
Keyword(s):  
Fluorescent Probe ◽  
Protein Interactions ◽  
Cytochromes P450 ◽  
Substrate Binding ◽  
Protein Protein Interactions ◽  
Heme Domain

SPR Analysis of Protein-Protein Interactions Involving Cytochromes P450 and Cytochrome b5 Integrated into Lipid Membrane

2020 ◽  
Vol 14 (2) ◽  
pp. 168-173
Author(s):  
L. A. Kaluzhskiy ◽  
P. V. Ershov ◽  
K. S. Kurpedinov ◽  
D. S. Sonina ◽  
E. O. Yablokov ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Lipid Membrane ◽  
Cytochromes P450 ◽  
Cytochrome B5 ◽  
Protein Protein Interactions

scholarly journals Protein-protein interactions of cytochromes P450 3A4 and 3A5 with their intermediate redox partners cytochromes b5

2015 ◽  
Vol 61 (4) ◽  
pp. 468-474
Author(s):  
O.V. Gnedenko ◽  
A.S. Ivanov ◽  
E.O. Yablokov ◽  
S.A. Usanov ◽  
D.V. Mukha ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Dissociation Constants ◽  
Cytochromes P450 ◽  
Cytochrome B5 ◽  
Electrochemical Analysis ◽  
Monooxygenase System ◽  
Protein Protein Interactions ◽  
Reduction Potentials ◽  
Drug Biotransformation ◽  
Redox Partners

Molecular interactions between proteins redox partners (cytochromes Р450 3А4, 3А5 and cytochrome b5) within the monooxygenase system, which is known to be involved in drug biotransformation, were investigated. Human cytochromes Р450 3A4 and 3А5 (CYP3A4 and CYP3A5) form complexes with various cytochromes b5: the microsomal (b5mc) and mitochondrial (b5om) forms of this protein, as well as with 2 “chimeric” proteins, b5(om-mc), b5(mc-om). Kinetic constants and equilibrium dissociation constants were determined by the SPR biosensor. Essential distinction between CYP3A4 and CYP3A5 was only observed upon their interactions with cytochrome b5om. Electroanalytical characteristics of electrodes with immobilized hemoproteins were obtained. The electrochemical analysis of CYP3A4, CYP3A5, b5mc, b5om, b5(om-mc), and b5(mc-om) immobilized on screen printed graphite electrodes modified with membranous matrix revealed that these proteins have very close reduction potentials -0.435  -0.350 V (vs. Ag/AgCl). Cytochrome b5mc was shown to be capable of stimulating the electrocatalytic activity of CYP3A4 in the presence of its substrate testosterone.

scholarly journals ‘Candidatus Liberibacter asiaticus’ Multimeric LotP Mediates Citrus sinensis Defense Response Activation

2021 ◽  
Vol 12 ◽  
Author(s):  
Marcelo L. Merli ◽  
Kaylie A. Padgett-Pagliai ◽  
Alexandra E. Cuaycal ◽  
Lucila Garcia ◽  
Maria Rosa Marano ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Plant Tissue ◽  
Substrate Binding ◽  
Lon Protease ◽  
Plant Proteins ◽  
Protein Protein Interactions ◽  
Citrus Plant ◽  
Protein Protein Interaction ◽  
Citrus Greening Disease ◽  
Liberibacter Asiaticus

‘Candidatus Liberibacter asiaticus’ is known as the most pathogenic organism associated with citrus greening disease. Since its publicized emergence in Florida in 2005, ‘Ca. L. asiaticus’ remains unculturable. Currently, a limited number of potential disease effectors have been identified through in silico analysis. Therefore, these potential effectors remain poorly characterized and do not fully explain the complexity of symptoms observed in citrus trees infected with ‘Ca. L. asiaticus.’ LotP has been identified as a potential effector and have been partially characterized. This protein retains structural homology to the substrate binding domain of the Lon protease. LotP interacts with chaperones like GroEL, Hsp40, DnaJ, and ClpX and may exercise its biological role through interactions with different proteins involved in proteostasis networks. Here, we evaluate the interactome of LotP—revealing a new protein–protein interaction target (Lon-serine protease) and its effect on citrus plant tissue integrity. We found that via protein–protein interactions, LotP can enhance Lon protease activity, increasing the degradation rate of its specific targets. Infiltration of purified LotP strained citrus plant tissue causing photoinhibition and chlorosis after several days. Proteomics analysis of LotP tissues recovering after the infiltration revealed a large abundance of plant proteins associated with the stabilization and processing of mRNA transcripts, a subset of important transcription factors; and pathways associated with innate plant defense were highly expressed. Furthermore, interactions and substrate binding module of LotP suggest potential interactions with plant proteins, most likely proteases.

Protein-protein interactions of cytochromes P450 3A4 and 3A5 with their intermediate redox partners cytochromes

2014 ◽  
Vol 60 (1) ◽  
pp. 17-27 ◽  
Author(s):  
O.V. Gnedenko ◽  
A.S. Ivanov ◽  
E.O. Yablokov ◽  
S.A. Usanov ◽  
D.V. Mukha ◽  
...  
Keyword(s):  
Cytochrome B ◽  
Protein Interactions ◽  
Dissociation Constants ◽  
Cytochromes P450 ◽  
Electrochemical Analysis ◽  
Monooxygenase System ◽  
Protein Protein Interactions ◽  
Reduction Potentials ◽  
Drug Biotransformation ◽  
Redox Partners

Molecular interactions between proteins redox partners (cytochromes Р450 3А4, 3А5 and cytochrome b ) within the monooxygenase system, which is known to be involved in drug biotransformation, were investigated. Human cytochromes Р450 3А4 and 3А5 (CYP3A4 and CYP3A5) form complexes with various cytochromes b : the microsomal ( b5mc ) and mitochondrial ( b5om ) forms of this protein, as well as with 2 “chimeric” proteins, b5(om-mc) , b5(mc-om) . Kinetic constants and equilibrium dissociation constants were determined by the SPR biosensor. Essential distinction between CYP3A4 and CYP3A5 was only observed upon their interactions with cytochrome b5om . Electroanalytical characteristics of electrodes with immobilized hemoproteins were obtained. The electrochemical analysis of CYP3A4, CYP3A5, b5mc, b5om , b5(om-mc) , and b5(mc-om) immobilized on screen printed graphite electrodes modified with membranous matrix revealed that these proteins have very close reduction potentials -0.435- -0.350 V (vs. Ag/AgCl). Cytochrome b mc was shown to be capable of stimulating the electrocatalytic activity of CYP3A4 to testosterone.

scholarly journals Protein-protein interactions in microsomal cytochrome P-450 isozyme LM2 and their effect on substrate binding

1989 ◽  
Vol 186 (1-2) ◽  
pp. 383-388 ◽  
Author(s):  
Peter HILDEBRANDT ◽  
Horacio GARDA ◽  
Anton STIER ◽  
Galina I. BACHMANOVA ◽  
Irina P. KANAEVA ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Substrate Binding ◽  
Protein Protein Interactions ◽  
Microsomal Cytochrome ◽  
Cytochrome P 450

scholarly journals The effect of isatin on protein-protein interactions between cytochrome b5 and cytochromes P450

2017 ◽  
Vol 63 (2) ◽  
pp. 170-175 ◽  
Author(s):  
P.V. Ershov ◽  
E.O. Yablokov ◽  
Yu.V. Mezentsev ◽  
L.A. Kalushskiy ◽  
A.V. Florinskaya ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Contact Region ◽  
Cytochromes P450 ◽  
Cytochrome B5 ◽  
Optical Biosensor ◽  
Test System ◽  
Feedback Mechanism ◽  
Dissociation Rate ◽  
Protein Protein Interactions ◽  
High Concentration

Cytochromes P450 (CYP) are involved in numerous biochemical processes including metabolism of xenobiotics, biosynthesis of cholesterol, steroid hormones etc. Since some CYP catalyze indol oxidation to isatin, we have hypothesized that isatin can regulate protein-protein interactions (PPI) between components of the CYP system thus representing a (negative?) feedback mechanism. The aim of this study was to investigate a possible effect of isatin on interaction of human CYP with cytochrome b5 (CYB5A). Using the optical biosensor test system employing surface plasmon resonance (SPR) we have investigated interaction of immobilized CYB5A with various CYP in the absence and in the presence of isatin. The SPR-based experiments have shown that a high concentration of isatin (270 mM) increases Kd values for complexes CYB5A/CYP3А5 and CYB5A/CYP3A4 (twofold and threefold, respectively), but has no influence on complex formation between CYB5A and other CYP (including indol-metabolizing CYP2C19 and CYP2E1). Isatin injection to the optical biosensor chip with the preformed molecular complex CYB5A/CYP3A4 caused a 30%-increase in its dissociation rate. Molecular docking manipulations have shown that isatin can influence interaction of CYP3А5 or CYP3A4 with CYB5A acting at the contact region of CYB5A/CYP.

Protein-Protein Interactions between Rat Hepatic Cytochromes P450 (P450s) and UDP-Glucuronosyltransferases (UGTs): Evidence for the Functionally Active UGT in P450-UGT Complex

2007 ◽  
Vol 22 (5) ◽  
pp. 367-376 ◽  
Author(s):  
Yuji Ishii ◽  
Megumi Iwanaga ◽  
Yoshio Nishimura ◽  
Shuso Takeda ◽  
Shin-ichi Ikushiro ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Cytochromes P450 ◽  
Protein Protein Interactions

scholarly journals CYP2D6-CYP2C9 Protein-Protein Interactions and Isoform-Selective Effects on Substrate Binding and Catalysis

2009 ◽  
Vol 37 (8) ◽  
pp. 1682-1689 ◽  
Author(s):  
Murali Subramanian ◽  
Michael Low ◽  
Charles W. Locuson ◽  
Timothy S. Tracy
Keyword(s):  
Protein Interactions ◽  
Substrate Binding ◽  
Protein Protein Interactions ◽  
Selective Effects

scholarly journals Study specificity of isatin interactions with P450 cytochromes

2018 ◽  
Vol 64 (1) ◽  
pp. 61-65 ◽  
Author(s):  
P.V. Ershov ◽  
Yu.V. Mezentsev ◽  
E.O. Yablokov ◽  
L.A. Kalushskiy ◽  
A.V. Florinskaya ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Protein Interactions ◽  
Cytochromes P450 ◽  
Cytochrome B5 ◽  
Direct Interaction ◽  
Protein Protein Interactions ◽  
Living Organisms ◽  
Adrenodoxin Reductase ◽  
Equilibrium Dissociation ◽  
Dissociation Constant Kd

Cytochrome P450-dependent monooxygenase systems exist basically in all living organisms, where they perform various important functions. The coordinated functioning of these systems involves many proteins participating in different protein-protein interactions (PPI). Previously, we have found that the endogenous non-peptide bioregulator isatin (indoledione-2,3), synthesized from indole by means of certain cytochromes P450 (e.g. P450 2E1, P450 2C19, P450 2A6) regulates affinity of some PPI. In this work, an attempt has been undertaken to register a direct interaction of isatin with a set of different proteins related to the functioning of cytochrome P450-dependent monooxygenase: five isoforms of cytochromes P450, two isoforms of cytochrome b5, cytochrome P450 reductase, adrenodoxin, adrenodoxin reductase and ferrochelatase. The study has shown that isatin binds specifically only to cytochromes P450 with high affinity (the equilibrium dissociation constant (Kd) is about 10-8 M).

Sign in / Sign up

Export Citation Format

Share Document