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. 2023 May 12;14(1):2735.
doi: 10.1038/s41467-023-37893-z.

Addendum: Sensing the allosteric force

Brankica Jankovic  1 , Olga Bozovic  1 , Peter Hamm  2
Affiliations

Addendum: Sensing the allosteric force

Brankica Jankovic et al. Nat Commun. .
No abstract available

PubMed Disclaimer

Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1. Fluorescence signal measured as a function of initial protein concentration.
Shown are parallel (red) and perpendicular (blue) polarizations of excitation light and fluorescence detection, respectively, exemplified here for the cis-state (filled circles, solid lines) and trans-state (open circles, dashed lines), respectively, at 30 °C. The peptide concentration has been 100 μM, and the excitation and detection wavelengths have been set to 280 and 360 nm, respectively, in order to maximize the fluorescence signal (it is actually not necessary to work at isosbestic points).
Fig. 2
Fig. 2. Fluorescence depolarisation and ITC measurements.
a Fluorescence anisotropy as a function of initial protein concentration and temperature with the photoswitch in the cis-state (red) or the trans-state (blue). peptide concentration has been 100 μM in all experiments. The anisotropy of the bound state is a bit larger than the theoretical upper limit of 0.4 due to slightly different detection sensitivities of the used fluorometer for both polarization directions. b ITC measurements of the trans-state. In either case, the data have been fit to a two-state binding equilibrium (solid lines).
Fig. 3
Fig. 3. Temperature dependence of ΔG, ΔH and −TΔS.
The binding free energy ΔG of the trans-state extracted from ITC (blue), along with the binding enthalpy ΔH (gray) and entropic contribution −TΔS (red). The binding free energies deduced from the fluorescence depolarization experiments (green) are shown for comparison as well.
Fig. 4
Fig. 4
Free energy landscape of the thermal cis-to-trans isomerization of the free protein P (red) or the ligand-bound protein PL (blue).
See this image and copyright information in PMC

Erratum for

  • doi: 10.1038/s41467-020-19689-7

References

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