Thermal shift assay of AtCinA.

<p>Melting temperature curves of purified enzyme (10 μg) were obtained in the presence of fluorescent probe SYPRO Orange in MilliQ^® water (◆), 160 mM sodium acetate pH 5.0 (●), 50 mM sodium phosphate pH 7.5 (▼), and 50 mM glycine pH 10.0 (■). Inset. Effect of different modulators on the melting temperature of AtCinA. Nucleotides were used at 1 mM (NAD⁺, FAD and NaMN), whereas hydroxy-ectoine (OH-Ect) and ammonium sulfate (AS) were used at 2 M and 1.5 M, respectively. Differences in Δ<i>Tm</i> were calculated by subtracting the MilliQ^® water <i>Tm</i> value from the <i>Tm</i> values obtained for the enzyme in the different conditions used.