RNA 5′ Polyphosphatase* is a Mg2+-independent phosphohydrolase discovered and characterized by Epicentre scientists. The enzyme sequentially removes the γ and β phosphates from 5′-triphosphorylated RNA (such as primary RNA transcripts):5′ pppN-OH 3′ → 5′ pN-OH 3′ + 2 PiRNAs with a 5′-diphosphorylated end are also converted to 5′-monophosphorylated RNA by RNA 5′ Polyphosphatase:5′ ppN-OH 3′ → 5′ pN-OH 3′ + PiRNA Polyphosphatase has no activity on RNA with a 5′ cap (e.g., 5′ m7GpppN-OH 3′), or a 5′-monophosphorylated end (5′ pN-OH 3′). However, both NTPs and dNTPs are substrates for the enzyme, yielding the corresponding NMPs and dNMPs + inorganic phosphate: (d)NTP → (d)NMP + 2Pi
Conversion of 5′-triphosphorylated RNA to 5′-monophosphorylated RNA for use in 5′-RNA ligation-tagging methods using T4 RNA Ligase. Analysis of 5′-end structure of RNA. Preparation of substrate RNA molecules for subsequent degradation using Epicentre"s Terminator? 5′-Phosphate-Dependent Exonuclease.
50% glycerol containing 0.05 M Tris-HCl (pH 7.5), 0.1 mM EDTA, 1 mM DTT, 0.1 M NaCl, and 0.1% Triton® X-100.
One unit of RNA 5′ Polyphosphatase releases 1 nmol of inorganic phosphate from ATP in 1 hour at 37 °C under standard assay conditions.