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Reverse-phase HPLC analysis of cAMP production. The elution profile of ATP, AMP, and cAMP (30 nmol) is shown in A, standard reactions lacking enzyme in B, reactions containing a control extract from BL21pLysS in C, and 1.0 μM rExoY in D. ATP and cAMP (30 nmol) treated with 3′:5′-cyclic nucleotide phosphodiesterase is shown in E. A reaction identical to that seen in D treated with 5 μg of 3′:5′-cyclic nucleotide phosphodiesterase for 10 min at 30°C is shown in F.
