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 Pi
RNAs with a 5´-diphosphorylated end are also converted to 5'-monophosphorylated RNA by RNA 5' Polyphosphatase:
5' ppN—OH 3' → 5' pN—OH 3' Pi
RNA 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
Unit Definition
One unit of RNA 5' Polyphosphatase releases 1 nmol of inorganic phosphate from ATP in 1 hour at 37°C under standard assay conditions.
Storage Buffer
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.
10X Reaction Buffer
0.5 M HEPES-KOH (pH 7.5), 1 M NaCl, 10 mM EDTA, 1% β-mercaptoethanol, and 0.1% Triton® X-100.
Inactivation/Inhibitors
RNA 5' Polyphosphatase is inhibited ~50% by 100 mM of inorganic phosphate (Pi) using ATP as a substrate.
Quality Control
RNA 5' Polyphosphatase is free of detectable DNA exo- and endonuclease, and RNase activities.
*Covered by issued and/or pending patents.
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