Reaction mechanism was concluded to involve 2'-OH nucleophilic attack by G53 on the phosphate bond connecting G53-U54.
[2] Under physiological pH and magnesium ion concentration, the rate constant of pistol ribozyme self-cleaving reaction was observed to be > 10 min−1.
Under optimum condition (pH = 7.0 - 9.0, and magnesium concentration above 50 mM), the rate constant detected to be > 100 min−1.
[2] Self-cleaving reactions were observed in the presence of 0.1 mM of various monovalent and divalent metal ions such as magnesium, manganese, calcium, cobalt, nickel, cadmium, barium, sodium, and lithium.
This implies that pistol ribozyme possess no specificity in the metal ion required in catalysis.