Protein dynamics and imidazole binding in cytochrome P450 enzymes

2006 ◽  
Vol 34 (6) ◽  
pp. 1170-1172 ◽  
Author(s):  
A. Verras ◽  
P.R. Ortiz de Montellano
Keyword(s):  
Cytochrome P450 ◽  
Retinoic Acid ◽  
Protein Dynamics ◽  
Active Sites ◽  
Triazole Ring ◽  
Crystallographic Analysis ◽  
Cytochrome P450 Enzymes ◽  
Endogenous Factors ◽  
Iron Inhibitors ◽  
Haem Iron

P450 (cytochrome P450) enzymes have major roles in the biosynthesis of endogenous factors such as steroids and eicosanoids, in the termination of the action of endogenous factors such as retinoic acid, in the metabolism of most drugs and xenobiotics and in the generation of toxic and carcinogenic products. Understanding the determinants of the substrate and inhibitor specificities of these enzymes is important for drug design. The crystallographic analysis of the deformability of two bacterial P450 active sites associated with the binding of azole (a class of inhibitors with an imidazole or triazole ring that co-ordinates to the haem iron) inhibitors described in the present study illustrates the importance of protein conformational malleability in the binding of imidazole derivatives.

scholarly journals MEMS directed evolution of two cytochrome P450 enzymes revealing distinct active-sites for convergent functions

2020 ◽  
Author(s):  
Li Ma ◽  
Fengwei Li ◽  
Xingwang Zhang ◽  
Hui Chen ◽  
Qian Huang ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Directed Evolution ◽  
Active Sites ◽  
Enzyme Engineering ◽  
Proof Of Concept ◽  
Cytochrome P450 Enzymes ◽  
Natural Evolution ◽  
One Pot ◽  
P450 Bm3 ◽  
Protein Enzyme

AbstractDirected evolution (DE) inspired by natural evolution (NE) has been achieving tremendous successes in protein/enzyme engineering. However, the conventional ‘one-protein-for-one-task’ DE cannot match the ‘multi-proteins-for-multi-tasks’ NE in terms of screening throughput and efficiency, thus often failing to meet the fast-growing demands for biocatalysts with desired properties. In this study, we design a novel ‘multi-enzyme-for-multi-substrate’ (MEMS) DE model and establish the proof-of-concept by running a NE-mimicking and higher-throughput screening on the basis of ‘two-P450s-against-seven-substrates’ (2P×7S) in one pot. With the significantly improved throughput and hit-rate, we witness a series of convergent evolution events of the two archetypal cytochrome P450 enzymes (P450 BM3 and P450cam) in laboratory. Further structural analysis of the two functionally convergent P450 variants provide important insights into how distinct active-sites can reach a common catalytic goal.

scholarly journals Bioactivity of curcumin on the cytochrome P450 enzymes of the steroidogenic pathway

2019 ◽  
Author(s):  
Patricia Rodríguez Castaño ◽  
Shaheena Parween ◽  
Amit V Pandey
Keyword(s):  
Cytochrome P450 ◽  
Active Sites ◽  
Curcuma Longa ◽  
Docking Studies ◽  
Steroid Metabolism ◽  
Dependent Manner ◽  
Cytochrome P450 Enzymes ◽  
Lead Compounds ◽  
Anti Cancer ◽  
Mild Reduction

AbstractTurmeric, a popular ingredient in the cuisine of many Asian countries, comes from the roots of theCurcuma longaand is known for its use in Chinese and Ayurvedic medicine. Turmeric is rich in curcuminoids, including curcumin, demethoxycurcumin, and bisdemethoxycurcumin. Curcuminoids have potent wound healing, anti-inflammatory, and anti-carcinogenic activities. While curcuminoids have been studied for many years, not much is known about their effects on steroid metabolism. Since many anti-cancer drugs target enzymes from the steroidogenic pathway, we tested the effect of curcuminoids on cytochrome P450 CYP17A1, CYP21A2, and CYP19A1 enzyme activities. When using 10 µg/ml of curcuminoids, both the 17α-hydroxylase as well as 17,20 lyase activities of CYP17A1 were reduced significantly. On the other hand, only a mild reduction in CYP21A2 activity was observed. Furthermore, CYP19A1 activity was also reduced up to ~20% of control when using 1-100 µg/ml of curcuminoids in a dose-dependent manner. Molecular docking studies confirmed that curcumin could dock into the active sites of CYP17A1, CYP19A1 as well as CYP21A2. In CYP17A1 and CYP19A1, curcumin docked within 2.5 Å of central heme while in CYP21A2 the distance from heme was 3.4 Å, which is still in the same range or lower than distances of bound steroid substrates. These studies suggest that curcuminoids may cause inhibition of steroid metabolism, especially at higher dosages. Also, the recent popularity of turmeric powder as a dilatory supplement needs further evaluation for the effect of curcuminoids on steroid metabolism. Molecular structure of curcuminoids could be modified to generate better lead compounds with inhibitory effects on CYP17A1 and CYP19A1 for potential drugs against prostate cancer and breast cancer.

scholarly journals Stereoselective Formation and Metabolism of 4-Hydroxy-Retinoic Acid Enantiomers by Cytochrome P450 Enzymes

2012 ◽  
Vol 287 (50) ◽  
pp. 42223-42232 ◽  
Author(s):  
Jakob A. Shimshoni ◽  
Arthur G. Roberts ◽  
Michele Scian ◽  
Ariel R. Topletz ◽  
Sean A. Blankert ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Retinoic Acid ◽  
Cytochrome P450 Enzymes

Phase I evaluation of all-trans retinoic acid with and without ketoconazole in adults with solid tumors.

1995 ◽  
Vol 13 (6) ◽  
pp. 1501-1508 ◽  
Author(s):  
J S Lee ◽  
R A Newman ◽  
S M Lippman ◽  
F V Fossella ◽  
M Calayag ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Retinoic Acid ◽  
Cytochrome P450 Enzymes ◽  
Short Term ◽  
Time Curve ◽  
Trans Retinoic Acid ◽  
All Trans Retinoic Acid ◽  
To Receive ◽  
Randomized Crossover Study ◽  
Over Time

PURPOSE All-trans retinoic acid (RA) induces accelerated plasma all-trans RA clearance, presumably via cytochrome P450 enzymes. This accelerated metabolism has been shown to be inhibited in the short term by the cytochrome P450 inhibitor ketoconazole. This study was conducted to evaluate the efficacy of ketoconazole in maintaining plasma all-trans RA levels over time. PATIENTS AND METHODS Using a randomized crossover study design, we randomly assigned six patients to receive all-trans RA (45 mg/m2 orally twice per day for 14 days of a 21-day cycle) for cycle 1 and the same dose of all-trans RA plus ketoconazole (400 mg orally for one dose, then 200 mg orally three times per day for 14 days) for cycle 2, and seven patients to receive the same treatment in the reverse order. Plasma all-trans RA levels were measured during the initial 8-hour period after all-trans RA ingestion on days 1 and 15 of cycles 1 and 2. RESULTS There was a marked decrease in plasma all-trans RA levels after 14 days of treatment, as measured by the area under the concentration-time curve (AUC), regardless of whether ketoconazole was given (from a baseline value of 857 to 44 ng/mL/h; P = .025) or not (from 1,355 to 308 ng/mL/h; P = .123). This lack of effect on plasma all-trans RA levels was not due to inadequate plasma ketoconazole levels. Ketoconazole administration was associated with more toxicity. No objective tumor responses were observed. CONCLUSION Ketoconazole does not appear to maintain adequate plasma all-trans RA levels over time.

scholarly journals Bioactivity of Curcumin on the Cytochrome P450 Enzymes of the Steroidogenic Pathway

2019 ◽  
Author(s):  
Patricia Rodríguez Castaño ◽  
Shaheena Parween ◽  
Amit V Pandey
Keyword(s):  
Breast Cancer ◽  
Prostate Cancer ◽  
Cytochrome P450 ◽  
Active Sites ◽  
Curcuma Longa ◽  
Steroid Metabolism ◽  
Dependent Manner ◽  
Cytochrome P450 Enzymes ◽  
Lead Discovery ◽  
Vitro Assay

Turmeric is a popular ingredient in the cuisine of many Asian countries. Turmeric is known for its use in Chinese and Ayurvedic medicine and comes from the roots of the Curcuma longa. Turmeric is rich in curcuminoids, including curcumin, demethoxycurcumin, and bisdemethoxycurcumin. Curcumin has potent anti-inflammatory and anti-carcinogenic activities. Since many anti-cancer drugs target enzymes from the steroidogenic pathway, we tested the bioactivity of curcuminoids on cytochrome P450 CYP17A1, CYP21A2, and CYP19A1 enzyme activities. Curcuminoids were extracted from turmeric with organic solvents. We conducted a cell-based assay for CYP17A1 and CYP21A2 activities using human adrenal cell line NCI-H295R. 3H-pregnenolone was used for CYP17A1 assays, and 3H-17alpha-hydroxyprogesterone was used as a substrate for CYP21A2. Curcuminoids were added at different concentrations and incubated for 24h. Steroids were separated by thin layer chromatography and analyzed by phosphorimager analysis. For CYP19A1 activity, an in vitro assay using endoplasmic reticulum from JEG3 were used with 3H-androstenedione as the substrate. Curcuminoids were incubated for 1h, and the formation of 3H-water from the androstenedione breakdown was measured by scintillation counting. When using 10 µg/ml of curcuminoids, both the 17-hydroxylase as well as 17,20 lyase activities of CYP17A1 were reduced significantly. On the other hand, CYP21A2 activity was only reduced to ~50% control. Furthermore, CYP19A1 activity was reduced to ~20% of control when using 1-100 µg/ml of curcuminoids in a dose-dependent manner. No effect on the activity of 5alpha reductase for the conversion of androstenedione to androstanedione was observed. Molecular docking studies confirmed that curcumin could dock into the active sites of CYP17A1, CYP19A1 as well as CYP21A2. In CYP17A1 and CYP19A1, curcumin docked within 2.5 Å of central heme while in CYP21A2 the distance from heme was 3.4 Å, which is still in the same range or lower than distances of bound steroid substrates. These studies show that curcuminoids may potentially cause inhibition of steroid metabolism, especially at higher dosages. The activities of CYP17A1 and CYP19A1 were inhibited by curcuminoids, which indicate potential anti-carcinogenic effects in case of prostate cancer as well as breast cancer, which can be targeted by inhibition of steroidogenesis. Also, the recent popularity of turmeric powder/curcumin as a dilatory supplement needs further evaluation for the effect of curcuminoids on steroid metabolism. Curcuminoids present in curcumin may affect activities of multiple steroid metabolizing cytochrome P450 enzymes. Computational docking suggests curcumin binds into the active sites of steroid metabolizing P450s and may serve as a model for lead discovery. Molecular structure of curcuminoids could be modified to generate better lead compounds with inhibitory effects on CYP17A1 and CYP19A1 for potential drugs against prostate cancer and breast cancer.

scholarly journals The quasi-irreversible inactivation of cytochrome P450 enzymes by paroxetine: a computational approach

2020 ◽  
Vol 18 (17) ◽  
pp. 3334-3345
Author(s):  
Emadeldin M. Kamel ◽  
Al Mokhtar Lamsabhi
Keyword(s):  
Cytochrome P450 ◽  
Active Site ◽  
Active Sites ◽  
Heme Iron ◽  
Computational Approach ◽  
Cytochrome P450 Enzymes

The potency of paroxetine as a P450 inhibitor is mainly attributed to the availability of two active sites on its structure, its compatibility with P450's active site and the ease of its tight coordination to heme iron.

scholarly journals Bioactivity of Curcumin on the Cytochrome P450 Enzymes of the Steroidogenic Pathway

2019 ◽  
Vol 20 (18) ◽  
pp. 4606 ◽  
Author(s):  
Rodríguez Castaño ◽  
Parween ◽  
Pandey
Keyword(s):  
Cytochrome P450 ◽  
Active Sites ◽  
Curcuma Longa ◽  
Docking Studies ◽  
Steroid Metabolism ◽  
Dependent Manner ◽  
Cytochrome P450 Enzymes ◽  
Lead Compounds ◽  
Anti Cancer ◽  
Mild Reduction

Turmeric, a popular ingredient in the cuisine of many Asian countries, comes from the roots of the Curcuma longa and is known for its use in Chinese and Ayurvedic medicine. Turmeric is rich in curcuminoids, including curcumin, demethoxycurcumin, and bisdemethoxycurcumin. Curcuminoids have potent wound healing, anti-inflammatory, and anti-carcinogenic activities. While curcuminoids have been studied for many years, not much is known about their effects on steroid metabolism. Since many anti-cancer drugs target enzymes from the steroidogenic pathway, we tested the effect of curcuminoids on cytochrome P450 CYP17A1, CYP21A2, and CYP19A1 enzyme activities. When using 10 µg/ml of curcuminoids, both the 17α-hydroxylase as well as 17,20 lyase activities of CYP17A1 were reduced significantly. On the other hand, only a mild reduction in CYP21A2 activity was observed. Furthermore, CYP19A1 activity was also reduced up to ~20% of control when using 1–100 µg/ml of curcuminoids in a dose-dependent manner. Molecular docking studies confirmed that curcumin could dock onto the active sites of CYP17A1, CYP19A1, as well as CYP21A2. In CYP17A1 and CYP19A1, curcumin docked within 2.5 Å of central heme while in CYP21A2 the distance from heme was 3.4 Å, which is still in the same range or lower than distances of bound steroid substrates. These studies suggest that curcuminoids may cause inhibition of steroid metabolism, especially at higher dosages. Also, the recent popularity of turmeric powder as a dilatory supplement needs further evaluation for the effect of curcuminoids on steroid metabolism. The molecular structure of curcuminoids could be modified to generate better lead compounds with inhibitory effects on CYP17A1 and CYP19A1 for potential drugs against prostate cancer and breast cancer.

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