Non-Conserved Amino Acid Residues Modulate the Thermodynamics of Zn(II) Binding to Classical ββα Zinc Finger Domains
Abstract
:1. Introduction
2. Results and Discussion
2.1. Classical Molecular Dynamics Simulations
2.2. Steered Molecular Dynamics Simulations
2.3. Spectroscopic Characterization
2.4. Isothermal Titration Calorimetry
2.5. Comparison with Previous Reports and Biological Significance
3. Materials and Methods
3.1. Materials
3.2. Computational Studies
3.2.1. Molecular Dynamics (MD) Simulations
3.2.2. Steered Molecular Dynamics (SMD) Simulations
3.3. Zinc Finger Peptide Synthesis
3.4. Determination of pKa Values of ZF Thiols
3.5. Circular Dichroism
3.6. Co(II) Binding to ZF Peptides
3.7. Zinc Finger Competition with Chelators
3.8. Isothermal Titration Calorimetry (ITC)
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
CD | circular dichroism |
CCHH | zinc finger with two Cys and two His residues being in Zn(II) coordination sphere |
CN | contact number |
CP1 | zinc finger consensus peptide 1 |
FES | free energy surface |
ITC | isothermal titration calorimetry |
MD | molecular dynamics |
PC | principal component |
PCA | principal component analysis |
PDB | protein data bank |
RMSF | root-mean-square fluctuations |
SMD | steered molecular dynamics |
ZF | zinc finger domain |
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ZF Peptide | −logKd7.4 | Kd7.4 (fM) | ∆(−logKd7.4) |
---|---|---|---|
CP1-1991 | 14.49 ± 0.05 | 3.2 ± 0.4 | 0 |
CP1-2015 | 12.30 ± 0.02 | 501 ± 23 | 2.19 |
CP1-1991-β | 14.40 ± 0.04 | 4.0 ± 0.4 | 0.09 |
CP1-1991-α | 13.50 ± 0.05 | 32 ± 3 | 0.99 |
CP1-1991-K/S | 14.04 ± 0.05 | 9.1 ± 0.9 | 0.45 |
CP1-1991-D/S | 14.29 ± 0.04 | 5.1 ± 0.4 | 0.20 |
CP1-1991-K/S-D/S | 13.52 ± 0.04 | 30 ± 3 | 0.97 |
ZF Peptide | ΔG° | ΔΔG° | −TΔS° | ΔHITC | ΔH° | ΔHCysH | ΔHZn-pep | ΔHfolding | nH |
---|---|---|---|---|---|---|---|---|---|
(kcal/mol) | |||||||||
CP-1991 | −19.77 ± 0.05 | 0 | −6.71 | −20.87 ± 0.19 | −13.06 | 13.23 | −26.29 | −6.29 | 1.56 |
CP-2015 | −16.78 ± 0.02 | 2.99 | −6.20 | −18.83 ± 0.39 | −10.58 | 13.97 | −24.44 | −4.55 | 1.64 |
CP-1991-β | −19.64 ± 0.04 | 0.13 | −7.02 | −20.61 ± 0.40 | −12.62 | 13.53 | −26.15 | −6.15 | 1.59 |
CP-1991-α | −18.42 ± 0.05 | 1.35 | −7.31 | −19.53 ± 0.43 | −11.10 | 14.27 | −25.37 | −5.37 | 1.68 |
CP-1991-K/S | −19.15 ± 0.05 | 0.62 | −10.78 | −16.82 ± 0.20 | −8.37 | 14.31 | −22.68 | −2.68 | 1.68 |
CP-1991-D/S | −19.49 ± 0.04 | 0.28 | −7.89 | −20.24 ± 0.35 | −11.60 | 14.01 | −25.62 | −5.62 | 1.65 |
CP-1991-K/S-D/S | −18.44 ± 0.04 | 1.33 | −7.23 | −19.68 ± 0.47 | −11.21 | 14.34 | −25.55 | −5.55 | 1.69 |
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Kluska, K.; Chorążewska, A.; Peris-Díaz, M.D.; Adamczyk, J.; Krężel, A. Non-Conserved Amino Acid Residues Modulate the Thermodynamics of Zn(II) Binding to Classical ββα Zinc Finger Domains. Int. J. Mol. Sci. 2022, 23, 14602. https://doi.org/10.3390/ijms232314602
Kluska K, Chorążewska A, Peris-Díaz MD, Adamczyk J, Krężel A. Non-Conserved Amino Acid Residues Modulate the Thermodynamics of Zn(II) Binding to Classical ββα Zinc Finger Domains. International Journal of Molecular Sciences. 2022; 23(23):14602. https://doi.org/10.3390/ijms232314602
Chicago/Turabian StyleKluska, Katarzyna, Aleksandra Chorążewska, Manuel David Peris-Díaz, Justyna Adamczyk, and Artur Krężel. 2022. "Non-Conserved Amino Acid Residues Modulate the Thermodynamics of Zn(II) Binding to Classical ββα Zinc Finger Domains" International Journal of Molecular Sciences 23, no. 23: 14602. https://doi.org/10.3390/ijms232314602