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. 2024 Oct 23;15(11):e02310-24. doi: 10.1128/mbio.02310-24

Fig 7.

Absorbance at 365 nm with increasing iron concentrations, circular dichroism spectra comparing apo-Fpa and holo-Fpa, and molar heat capacity changes with temperature in two conditions are depicted.

Fpa is an Fe(II) binding protein. (A) Fe-binding affinity to Fpa measured using Mag-Fura-2 within a competition-based assay. Average titration spectral points of Fe(II) ions into Fpa with overall simulation in solid line with representative titration absorption spectra inset display. Mag-fura-2 to protein ratios were varied, and 1:1 data are shown. Spectra were collected in duplicate using independent samples to ensure spectral reproducibility. (B) Circular dichroism (CD) structural results for apo and Fe(II) loaded holo-Fpa. Representative CD spectra comparing apo- (black) and holo- (blue) Fpa. (C) Differential scanning calorimetric spectra for apo- and Fe(II) loaded holo-Fpa. Overall simulation of molar heat capacity vs temperature spectra for apo-Fpa (A) and holo-Fpa (B). Overall simulations are displayed in solid lines, with individual simulated features in gray and raw data shown in a black dashed line.