The Development of CDC25A-Derived Phosphoseryl Peptides That Bind 14-3-3ε with High Affinities
Abstract
:1. Introduction
2. Results
2.1. Computational Design of the Peptide Analogs
2.2. MD Simulations of 14-3-3ε–Peptide Complexes
2.3. Binding Free Energy of the 14-3-3ε–Peptide Complexes
2.4. Secondary Structure of the Peptide Analogs
2.5. Experimental Validation of Binding of the Peptides to 14-3-3ε
3. Discussion
4. Materials and Methods
4.1. MD Simulations
4.1.1. 14-3-3ε–Peptide Complexes Preparation
4.1.2. MD Simulations of 14-3-3ε–Peptide Complex
4.1.3. Trajectory Analysis
4.1.4. Computational Ala Scanning and Free Energy Calculation Using the MM/PBSA Method
4.1.5. SMD and US Simulations
4.2. Protein and Peptides
4.3. CD Spectrometry Measurements
4.4. Differential Scanning Fluorimetry (DSF)
4.5. Surface Plasmon Resonance (SPR)
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Peptide | Sequences * |
---|---|
pS | Ac-Thr-Gln-Arg-Gln-Asn-pSer-Ala-Pro-Ala-Arg-Met-Leu-Ser-Ser-NH2 |
pS(173−185) | Ac-Thr-Gln-Arg-Gln-Asn-pSer-Ala-Pro-Ala-Arg-Met-Leu-Ser-NH2 |
pS(173−184) | Ac-Thr-Gln-Arg-Gln-Asn-pSer-Ala-Pro-Ala-Arg-Met-Leu-NH2 |
pS(173−183) | Ac-Thr-Gln-Arg-Gln-Asn-pSer-Ala-Pro-Ala-Arg-Met-NH2 |
pS(174−183) | Ac-Gln-Arg-Gln-Asn-pSer-Ala-Pro-Ala-Arg-Met-NH2 |
pS(174–182) | Ac-Gln-Arg-Gln-Asn-pSer-Ala-Pro-Ala-Arg-NH2 |
Motifs | P−4 | P−3 | P−2 | P−1 | P | P+1 | P+2 | ||||
---|---|---|---|---|---|---|---|---|---|---|---|
Motif1 | Arg | Ser | Xaa | pSer/Thr | Xaa | Pro | |||||
Motif 2 | Arg | Xaa | Tyr/Phe | Xaa | pSer/Thr | Xaa | Pro | ||||
pS(174–182) | Ac | Gly | Arg | Gln | Asn | pSer | Ala | Pro | Ala | Arg | NH2 |
Peptide | Sequences * |
---|---|
pS | Ac-Thr-Gln-Arg-Gln-Asn-pSer-Ala-Pro-Ala-Arg-Met-Leu-Ser-Ser-NH2 |
pS(174–182) | Ac-Gln-Arg-Gln-Asn-pSer-Ala-Pro-Ala-Arg-NH2 |
[Phe176]pS(174–182) | Ac-Gln-Arg-Phe-Asn-pSer-Ala-Pro-Ala-Arg-NH2 |
[Tyr176]pS(174–182) | Ac-Gln-Arg-Tyr-Asn-pSer-Ala-Pro-Ala-Arg-NH2 |
[Lys177]pS(174–182) | Ac-Gln-Arg-Gln-Lys-pSer-Ala-Pro-Ala-Arg-NH2 |
[Phe177]pS(174–182) | Ac-Gln-Arg-Gln-Phe-pSer-Ala-Pro-Ala-Arg-NH2 |
[Trp177]pS(174–182) | Ac-Gln-Arg-Gln-Trp-pSer-Ala-Pro-Ala-Arg-NH2 |
[Phe179]pS(174–182) | Ac-Gln-Arg-Gln-Asn-pSer-Phe-Pro-Ala-Arg-NH2 |
[Tyr179]pS(174–182) | Ac-Gln-Arg-Gln-Asn-pSer-Tyr-Pro-Ala-Arg-NH2 |
Peptide | ΔTm/°C | KD ± SD/μM | ΔG ± SD/kJ mol−1 |
---|---|---|---|
pS | 0.35 ± 0.16 | 0.22 ± 0.01 | −37.96 ± 0.09 |
pS(174–182) | 0 | 9.05 ± 0.83 | −28.76 ± 0.23 |
[Phe176]pS(174–182) | 1.08 ± 0.34 | 0.36 ± 0.63 | −36.48 ± 0.63 |
[Tyr176]pS(174–182) | 1.10 ± 0.32 | 0.41 ± 0.10 | −36.76 ± 0.46 |
[Lys177]pS(174–182) | 0.23 ± 0.09 | 5.23 ± 0.47 | −30.12 ± 0.23 |
[Phe179]pS(174–182) | 0 | 58.28 ± 4.67 | −24.15 ± 0.2 |
[Tyr179]pS(174–182) | 0 | 115.53 ± 16.7 | −22.47 ± 0.39 |
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Kamayirese, S.; Maity, S.; Hansen, L.A.; Lovas, S. The Development of CDC25A-Derived Phosphoseryl Peptides That Bind 14-3-3ε with High Affinities. Int. J. Mol. Sci. 2024, 25, 4918. https://doi.org/10.3390/ijms25094918
Kamayirese S, Maity S, Hansen LA, Lovas S. The Development of CDC25A-Derived Phosphoseryl Peptides That Bind 14-3-3ε with High Affinities. International Journal of Molecular Sciences. 2024; 25(9):4918. https://doi.org/10.3390/ijms25094918
Chicago/Turabian StyleKamayirese, Seraphine, Sibaprasad Maity, Laura A. Hansen, and Sándor Lovas. 2024. "The Development of CDC25A-Derived Phosphoseryl Peptides That Bind 14-3-3ε with High Affinities" International Journal of Molecular Sciences 25, no. 9: 4918. https://doi.org/10.3390/ijms25094918