Neurosteroids: Structure-Uptake Relationships and Computational Modeling of Organic Anion Transporting Polypeptides (OATP)1A2
Abstract
:1. Introduction
2. Results
2.1. Cellular Uptake Studies
2.1.1. Cellular Uptake of 3α5β-Pregnanolone (1, CAS Number 128-20-1)
2.1.2. Cellular Uptake of Neurosteroids with Ester Bond (2 and 3)
2.1.3. Cellular Uptake of Neurosteroids Having 3-Carbon Amide Residues (4 and 5)
2.1.4. Cellular Uptake of Neurosteroids Having Medium Length 4-Carbon Amide Residues (6, 7, and 8)
2.1.5. Cellular Uptake of Neurosteroids with Longer Length 5-Carbon Amide Residues (9, 10, and 11)
2.1.6. Transporter-Mediated Uptake of Neurosteroids 1–11 at pH 5.5
2.1.7. Transporter-Mediated Uptake of Neurosteroids 1–11 at pH 7.4
2.2. Molecular Modeling
2.2.1. Homology Model—OATP1A2 Model Shows Two Binding Sites
2.2.2. OATP1A2 Substrate E3S Displays Two Potential Binding Modes
2.2.3. Binding of Compounds 2 and 3
2.2.4. Binding of Compounds 4 and 5
2.2.5. Binding of Compounds 6, 7, and 8
2.2.6. Binding of Compounds 9, 10, and 11
2.2.7. Principal Component Analysis (PCA)
3. Discussion
4. Materials and Methods
4.1. Chemicals
4.2. Cell Cultures
4.3. Cellular Uptake of Neurosteroids
4.4. High-Performance Liquid Chromatography (HPLC) Analyses
4.5. Data Analysis
4.6. Molecular Modeling
4.6.1. Homology Model Generation and Protein Preparation
4.6.2. Binding Site Prediction
4.6.3. Ligand Preparation and Docking
4.6.4. Molecular Dynamics (MD) Simulations
4.6.5. Principal Component Analysis (PCA)
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Sample Availability
References
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Transport Mechanism Type 1 | Transport Mechanism Type 2 | |||||
---|---|---|---|---|---|---|
Vmax (pmol/mg/min) | Km (µM) | Vmax/Km | Vmax (pmol/mg/min) | Km (µM) | Vmax/Km | |
1 | 26.3 ± 4.2 | 55.4 | 0.47 | - | - | - |
2 | n.d. a | n.d. a | n.d. a | |||
3 | 3.0 ± 0.4 | 7.7 | 0.39 | 8.2 ± 0.1 | 25.2 | 0.33 |
4 | 13.9 ± 6.7 | 232.8 | 0.059 | |||
5 | 5.3 ± 0.4 | 76.4 | 0.07 | 34.7 ± 7.7 | 959.6 | 0.04 |
6 | 3.9 ± 0.5 | 13.8 | 0.28 | 10.2 ± 0.1 | 91.26 | 0.11 |
7 | 3.0 ± 0.1 | 3.0 | 1.0 | 23.0 ± 0.5 | 127.4 | 0.18 |
8 | 8.0 ± 2.1 | 10.9 | 0.73 | 28.2 ± 0.1 | 42.8 | 0.66 |
9 | 5.5 ± 0.6 | 9.2 | 0.60 | 26.5 ± 3.0 | 67.8 | 0.39 |
10 | 15.4 ± 6.5 | 23.0 | 0.67 | 150.4 ± 9.9 | 281.6 | 0.53 |
11 | 17.2 ± 2.1 | 24.2 | 0.71 | - | - | - |
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Adla, S.K.; Tonduru, A.K.; Kronenberger, T.; Kudova, E.; Poso, A.; Huttunen, K.M. Neurosteroids: Structure-Uptake Relationships and Computational Modeling of Organic Anion Transporting Polypeptides (OATP)1A2. Molecules 2021, 26, 5662. https://doi.org/10.3390/molecules26185662
Adla SK, Tonduru AK, Kronenberger T, Kudova E, Poso A, Huttunen KM. Neurosteroids: Structure-Uptake Relationships and Computational Modeling of Organic Anion Transporting Polypeptides (OATP)1A2. Molecules. 2021; 26(18):5662. https://doi.org/10.3390/molecules26185662
Chicago/Turabian StyleAdla, Santosh Kumar, Arun Kumar Tonduru, Thales Kronenberger, Eva Kudova, Antti Poso, and Kristiina M. Huttunen. 2021. "Neurosteroids: Structure-Uptake Relationships and Computational Modeling of Organic Anion Transporting Polypeptides (OATP)1A2" Molecules 26, no. 18: 5662. https://doi.org/10.3390/molecules26185662