The fluorescence spectra were collected at 25°C using the PTI QuantaMaster 400 (Horiba, Canada) with monochromators for both excitation and emission. Fluorescence spectra of MMOH [0.32 μM MMOH in 300 μl of 25 mM Mops (pH 7.0) and 1 mM DTT] were determined using an excitation wavelength of 282 nm. Fluorescence was quantified by the integration of fluorescence emission bands with a maximum at 363 nm. The MMOH fluorescence was quenched by titration with MMOB, MMOD, and truncated MMODs (residues 1 to 74, 12 to 111, and 12 to 74). These proteins were titrated against MMOH in the concentration range from 0 to 15 μM. Dissociation constants were determined by fitting the intensity of emission at 363 nm (Origin 2018b and Wolfram Mathematica). Independent experiments were repeated in triplicate to allow the calculation of the average ± SEM.

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