Role of Long-Range Protein Dynamics in Different Thymidylate Synthase Catalyzed Reactions
Abstract
:1. Introduction
2. Results and Discussion
2.1. Binding Mechanism of Y209W Mutant
2.2. Temperature Dependence of Intrinsic KIEs in the Y209W Mutant
Proton Abstraction | Hydride Transfer | |||
---|---|---|---|---|
WT a | Y209W | WT b | Y209W c | |
AH/AT | 8.3 (±1.0) ×10−6 | 1.4 (±0.1) ×10−6 | 6.8 (±2.8) | 3.6 (±0.9) |
∆Ea H–T (kcal/mol) | −8.0 (±0.1) | −7.70 (±0.40) | −0.02 (±0.25) | −0.5 (±0.1) |
2.3. Effect of Y209W Mutation on Catalytic Steps Other than the Two C–H Activations
3. Experimental Section
3.1. Materials
3.2. Synthesis of [2-14C,5-2H] dUMP(>99.5% D)
3.3. Competitive KIEs on the Proton Abstraction (Step 4 in Scheme 1)
4. Conclusions
Supplementary Files
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Abeysinghe, T.; Kohen, A. Role of Long-Range Protein Dynamics in Different Thymidylate Synthase Catalyzed Reactions. Int. J. Mol. Sci. 2015, 16, 7304-7319. https://doi.org/10.3390/ijms16047304
Abeysinghe T, Kohen A. Role of Long-Range Protein Dynamics in Different Thymidylate Synthase Catalyzed Reactions. International Journal of Molecular Sciences. 2015; 16(4):7304-7319. https://doi.org/10.3390/ijms16047304
Chicago/Turabian StyleAbeysinghe, Thelma, and Amnon Kohen. 2015. "Role of Long-Range Protein Dynamics in Different Thymidylate Synthase Catalyzed Reactions" International Journal of Molecular Sciences 16, no. 4: 7304-7319. https://doi.org/10.3390/ijms16047304