SPR and electrochemical analyses of interactions between CYP3A4 or 3A5 and cytochrome b5

Gnedenko, O. V.; Yablokov, E. O.; Usanov, Sergey A.; Mukha, Dzmitry; Sergeev, G. V.; Булко, Т. В.; Kuzikov, Alexey V.; Moskaleva, Natalia E.; Shumyantseva, Victoria V.; Ivanov, A. S.; Archakov, Alexander I.

doi:10.1016/j.cplett.2013.12.041

Cited by 27 publications

(9 citation statements)

References 34 publications

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“…The calculated K d values of TBXAS1 • CYP11B2 and TBXAS1 • CYP2E1 complex formation were (6.9 ± 0.3) × 10 -7 M and (4.3 ± 0.4) × 10 -7 M, respectively. These values are comparable to K d of the complexes of various cytochromes P450 with their functional partners (CPR, CYB5A, ADX) [ 23 , 34 - 37 ]. It is important to note that, while the difference in the K d values of complex formation is about twofold, TBXAS1 • CYP11B2 and TBXAS1 • CYP2E1 interactions are very different in their kinetic parameters.…”

Section: Resultssupporting

confidence: 58%

Direct Molecular Fishing of New Protein Partners for Human Thromboxane Synthase

Svirid¹,

Ershov²,

Yablokov³

et al. 2017

Acta Naturae

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Thromboxane synthase (TBXAS1) catalyzes the isomerization reaction of prostaglandin H 2 producing thromboxane A 2 , the autocrine and paracrine factor in many cell types. A high activity and metastability by these arachidonic acid derivatives suggests the existence of supramolecular structures that are involved in the regulation of the biosynthesis and directed translocation of thromboxane to the receptor. The objective of this study was to identify TBXAS1 protein partners from human liver tissue lysate using a complex approach based on the direct molecular fishing technique, LC-MS/MS protein identification, and protein-protein interaction validation by surface plasmon resonance (SPR). As a result, 12 potential TBXAS1 protein partners were identified, including the components regulating cytoskeleton organization (BBIP1 and ANKMY1), components of the coagulation cascade of human blood (SERPINA1, SERPINA3, APOH, FGA, and FN1), and the enzyme involved in the metabolism of xenobiotics and endogenous bioregulators (CYP2E1). SPR validation on the Biacore 3000 biosensor confirmed the effectiveness of the interaction between CYP2E1 (the enzyme that converts prostaglandin H 2 to 12-HHT/thromboxane A 2 proantagonist) and TBXAS1 (K d = (4.3 ± 0.4) × 10 -7 M). Importantly, the TBXAS1•CYP2E1 complex formation increases fivefold in the presence of isatin (indole-2,3-dione, a low-molecular nonpeptide endogenous bioregulator, a product of CYP2E1). These results suggest that the interaction between these hemoproteins is important in the regulation of the biosynthesis of eicosanoids. KEYWORDS Thromboxane synthase (CYP5A1, TBXAS1), cytochrome P450, surface plasmon resonance, direct molecular fishing, protein partners, isatin. ABBREVIATIONS Protein-protein interaction (PPI), surface plasmon resonance (SPR), association rate constant (k on ), dissociation rate constant (k off ), equilibrium dissociation constant (K d ).

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Section: Resultssupporting

confidence: 58%

Direct Molecular Fishing of New Protein Partners for Human Thromboxane Synthase

Svirid¹,

Ershov²,

Yablokov³

et al. 2017

Acta Naturae

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“…У человека обнаружено более 50 типов CYP [1], участвующих в разнообразных биохимических и физиологических процессах, таких как метаболизм ксенобиотиков (в том числе лекарственных соединений) (CYP3A4, CYP3A5, CYP2C9 и CYP1B1 [2][3][4][5]), биосинтез холестерина и стероидных гормонов (CYP11A1 [6], CYP17A1 [7], CYP11B1 [8]). Небольшой гемопротеин цитохром b 5 (СYB5A) взаимодействует с рядом CYP, выполняя в этих монооксигеназных системах роль переносчика электронов и аллостерического эффектора [9][10][11][12][13][14]. Известно, что связывание субстрата CYP способно изменять сродство комплексов в комплексах CYP/CYB5A [15].…”

Section: Introductionunclassified

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

et al. 2017

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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.

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“…These values are comparable to K d of the complexes of various cytochromes P450 with their functional partners (CPR, CYB5A, ADX) [23, 34-37]. It is important to note that, while the difference in the K d values of complex formation is about twofold, TBXAS1 • CYP11B2 and TBXAS1 • CYP2E1 interactions are very different in their kinetic parameters.…”

Section: Resultsmentioning

confidence: 53%

Direct Molecular Fishing of New Protein Partners for Human Thromboxane Synthase

Свирид¹,

Ershov²,

Yablokov³

et al. 2017

Acta Naturae

View full text Add to dashboard Cite

Thromboxane synthase (TBXAS1) catalyzes the isomerization reaction of prostaglandin H2 producing thromboxane A2, the autocrine and paracrine factor in many cell types. A high activity and metastability by these arachidonic acid derivatives suggests the existence of supramolecular structures that are involved in the regulation of the biosynthesis and directed translocation of thromboxane to the receptor. The objective of this study was to identify TBXAS1 protein partners from human liver tissue lysate using a complex approach based on the direct molecular fishing technique, LC-MS/MS protein identification, and protein-protein interaction validation by surface plasmon resonance (SPR). As a result, 12 potential TBXAS1 protein partners were identified, including the components regulating cytoskeleton organization (BBIP1 and ANKMY1), components of the coagulation cascade of human blood (SERPINA1, SERPINA3, APOH, FGA, and FN1), and the enzyme involved in the metabolism of xenobiotics and endogenous bioregulators (CYP2E1). SPR validation on the Biacore 3000 biosensor confirmed the effectiveness of the interaction between CYP2E1 (the enzyme that converts prostaglandin H2 to 12-HHT/thromboxane A2 proantagonist) and TBXAS1 (Kd = (4.3 ± 0.4) × 10-7 M). Importantly, the TBXAS1•CYP2E1 complex formation increases fivefold in the presence of isatin (indole-2,3-dione, a low-molecular nonpeptide endogenous bioregulator, a product of CYP2E1). These results suggest that the interaction between these hemoproteins is important in the regulation of the biosynthesis of eicosanoids.

show abstract

SPR and electrochemical analyses of interactions between CYP3A4 or 3A5 and cytochrome b5

Cited by 27 publications

References 34 publications

Direct Molecular Fishing of New Protein Partners for Human Thromboxane Synthase

Direct Molecular Fishing of New Protein Partners for Human Thromboxane Synthase

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

Direct Molecular Fishing of New Protein Partners for Human Thromboxane Synthase

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