Structural Insight into Polymerase Mechanism via a Chiral Center Generated with a Single Selenium Atom
Abstract
:1. Introduction
2. Results
2.1. Overall Structures of the Complexes
2.2. The Absolute Configurations of the Two dGTPαSe Diastereomers
2.3. Configuration Inversion at the α-Phosphorus Center in the Polymerization Reaction
2.4. The In-Line SN2 Attacking Observed in One Single Crystal with dGTPαSe-Rp
2.5. DNA Polymerase Recognizing dNTPαSe as the Substrates in Thermodynamic and Kinetic Studies
3. Discussion
4. Materials and Methods
4.1. Protein Expression and Purification
4.2. Crystallization and X-ray-Diffraction Data Collection
4.3. Structure Determination and Refinement
4.4. dNTP Binding Assay In Vitro
4.5. The Apparent and Standard Kinetic Study of Polymerase
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Qin, T.; Hu, B.; Zhao, Q.; Wang, Y.; Wang, S.; Luo, D.; Lyu, J.; Chen, Y.; Gan, J.; Huang, Z. Structural Insight into Polymerase Mechanism via a Chiral Center Generated with a Single Selenium Atom. Int. J. Mol. Sci. 2023, 24, 15758. https://doi.org/10.3390/ijms242115758
Qin T, Hu B, Zhao Q, Wang Y, Wang S, Luo D, Lyu J, Chen Y, Gan J, Huang Z. Structural Insight into Polymerase Mechanism via a Chiral Center Generated with a Single Selenium Atom. International Journal of Molecular Sciences. 2023; 24(21):15758. https://doi.org/10.3390/ijms242115758
Chicago/Turabian StyleQin, Tong, Bei Hu, Qianwei Zhao, Yali Wang, Shaoxin Wang, Danyan Luo, Jiazhen Lyu, Yiqing Chen, Jianhua Gan, and Zhen Huang. 2023. "Structural Insight into Polymerase Mechanism via a Chiral Center Generated with a Single Selenium Atom" International Journal of Molecular Sciences 24, no. 21: 15758. https://doi.org/10.3390/ijms242115758