P1, P3‐bis(5′‐adenosyl)triphosphate (Ap3A) as a substrate and a product of mammalian tryptophanyl‐tRNA synthetase

Bovine tryptophanyl-tRNA synthetase (TrpRS, E.C.6.1.1.2) is unable to catalyze in vitro formation of Ap4A in contrast to some other aminoacyl-tRNA synthetases. However, in the presence of L-tryptophan, ATP-Mg2+ and ADP the enzyme catalyzes the Ap3A synthesis via adenylate intermediate. Ap3A (not Ap4A) may serve as a substrate for TrpRS in the reaction of E.(Trp approximately AMP) formation and in the tRNA(Trp) charging. The Km value for Ap3A was higher than the Km for ATP (approx. 1.00 vs. 0.22 mM) and Vmax was 3 times lower than for ATP. The Zn(2+)-deficient enzyme catalyzes Ap3A synthesis in the absence of exogenous ADP due to ATPase activity of Zn(2+)-deprived TrpRS. The inability of mammalian TrpRS to synthesize Ap4A, might be considered as a molecular tool preventing the removal of Zn2+ due to chelation by Ap4A and therefore preserving the enzyme activity.