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. 2024 Jun 6;15(7):e01220-24. doi: 10.1128/mbio.01220-24

Fig 2.

Phos-tag gels for BT4124 variants and time points; quantification of phosphorylated protein levels; data on initial rates of BT4124 protein phosphorylation; histogram comparing efficiency of autophosphorylation between RR and REC domains of four HTCSs.

Isolated REC domains autophosphorylate faster than HTCS RRs with linked DNA binding domains. (A–C) Autophosphorylation kinetics of BT4124. Phosphorylation of BT4124 proteins was analyzed using Phos-tag gels at indicated times after addition of 20 mM PAM (A). One representative example is shown for each protein. Fractions of phosphorylated proteins from panel A were quantified to derive the exponential trendline of phosphorylation in panel B. Initial rates of phosphorylation were calculated from early stages of reaction (inset in panel B). (C) Kinetic characterization of BT4124 phosphorylation. Independent measurements of initial rates are plotted as circles. Solid lines indicate individual fits with the Michaelis-Menten equation and the shaded areas represent the 95% confidence intervals. (D) Comparison of phosphorylation efficiency between RRs and REC domains across different HTCS proteins. Error bars indicate the standard deviation calculated as described in Materials and Methods.