Determination of intracellular protein–ligand binding affinity by competition binding in-cell NMR

Luchinat, E.; Barbieri, L.; Cremonini, M.; Pennestri, M.; Nocentini, A.; Supuran, C.T.; Banci, L.

doi:10.1107/S2059798321009037

Acta Crystallographica Section D
Acta Crystallographica
Section D STRUCTURAL BIOLOGY

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Figure 5
Bioreactor in-cell NMR of cells treated with constant 10 µM MZA and increasing amounts of ETZ (bioreactor run 2 in Table 1

) and subsequent data analysis. (a–c) Representative ¹H–¹⁵N NMR spectra at different time points and concentrations of ETZ. Signals arising from ETZ are marked with magenta arrows. (d) NMR spectra of the pure components, i.e. CA II–MZA (black) and CA II–ETZ (magenta), reconstructed by MCR-ALS. (e) Concentration profiles of CA II–MZA (black squares) and CA II–ETZ (magenta circles) obtained by MCR-ALS. (f) Bound fractions obtained from the plateau values after each step of the run plotted as a function of ETZ concentration. Binding curves from nonlinear fitting are shown as dashed lines.

STRUCTURAL
BIOLOGY

ISSN: 2059-7983

Volume 77| Part 10| October 2021| Pages 1270-1281

https://doi.org/10.1107/S2059798321009037

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