ZFIN Image: Samper-Martín et al., 2021, Fig. 2

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Figure Caption

Fig. 2

Figure 2. Nudt3 substrate specificity is ion dependent

(A) Divalent ion dependence of Nudt3 endo-polyPase activity. Quantification of DAPI-stained PAGE-urea gels as in Figure 1. Nudt3 (100 ng) and polyP₁₀₀ were incubated for 1 h in the presence of the mentioned ions (5 mM each), EDTA, or EGTA (10 mM). 100 ng of yeast recombinant endo-polyPase Ddp1 (Nudt3 homolog) is used as control. Mean ± SEM of three independent experiments.

(B) Exo-polyPase activity of Nudt3. The enzymatic reaction was as in (A). The activity was measured as free phosphate production using the malachite green method. Yeast recombinant exo-polyPase Ppx1 (1 ng) is used as a control. Mean ± SEM of three independent experiments.

(C) Enzymatic activity quantification of Nudt3 on the noted substrates (up to down: polyP_100, 5-InsP₇, and Ap5A) in the presence of the indicated cations (5 mM). Mean ± SEM of three independent experiments. ∗∗∗p < 0.001; ∗∗p < 0.01; ∗p < 0.05.

Acknowledgments

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