Heat shock inducible lysyl-tRNA synthetase ofEscherichia coli(LysU) is known to be a highly efficient diadenosine 5¢,5¢¢¢-P 1 ,P 4 -tetraphosphate (Ap4A) synthase. However, we use an ion-exchange HPLC technique to demonstrate that active LysU mixtures actually have a dual catalytic activ-ity, initially producing Ap4 A from ATP, before converting that tetraphos-phate to a triphosphate.