(A) Kinetic of the rPpx1 polyP digestion. rPpx1 (10 ng) was
incubated with 250 ng of commercial polyP or yeast polyP, in 20 mM
Tris-HCl pH 7.50 containing 5 mM magnesium acetate and 100 mM ammonium
acetate at 37°C. Samples were taken at the indicated times to quantify
the released Pi. Mean ± SEM from 3 independent experiments is shown.
(B) rPpx1 activity on polyP in the presence of increasing
amount of DNA. rPpx1 (10 ng) was incubated with 100 ng of commercial
polyP in 20 mM Tris-HCl pH 7.50 containing 5 mM magnesium acetate and
100 mM ammonium acetate and at 37°C during 20 min with increasing
concentrations of DNA (both circular and linear). The graph represents
the released Pi in each condition. Mean ± SEM from 3 independent
experiments is shown.
(C) rPpx1 activity on polyP in the presence of increasing
amount of RNA. Same experiment as in B, but with increasing
concentrations of RNA. The graph represents the released Pi in each
condition. Mean ± SEM from 3 independent experiments is shown.
(D) Kinetics of rPpx1 polyP digestion in the presence of DNA
and RNA. rPpx1 (1 ng) was incubated with 250 ng of yeast polyP
previously treated or not with a DNAse/ RNAse solution in 20 mM Tris-HCl
pH 7.50 containing 5 mM magnesium acetate and 100 mM ammonium acetate at
37°C. Samples were taken at the indicated times and the released Pi was
quantified. Mean ± SEM from 3 independent experiments is shown.