Mechanisms of ATP to cAMP Conversion Catalyzed by the Mammalian Adenylyl Cyclase: A Role of Magnesium Coordination Shells and Proton Wires

We report a mechanism of adenosine triphosphate (ATP) to cyclic adenosine monophosphate (cAMP) conversion by the mammalian type V adenylyl cyclase revealed in molecular dynamics (MD) and quantum mechanics/molecular mechanics (QM/MM) simulations. We characterize a set of computationally derived enzyme-substrate (ES) structures showing an important role of coordination shells of magnesium ions in the solvent accessible active site. Several stable six-fold coordination shells of Mg_A²⁺ are observed in MD simulations of ES complexes. In the lowest energy ES conformation, the coordination shell of Mg_A²⁺ does not include the O_δ1 atom of the conserved Asp440 residue. Starting from this conformation, a one-step reaction mechanism is characterized which includes proton transfer from the ribose O^3'H^3' group in ATP to Asp440 via a shuttling water molecule and P^A-O^3A bond cleavage and O^3'-P^A bond formation. The energy profile of this route is consistent with the observed reaction kinetics. In a higher energy ES conformation, Mg_A²⁺ is bound to the O_δ1(Asp440) atom as suggested in the relevant crystal structure of the protein with a substrate analog. The computed energy profile initiated by this ES is characterized by higher energy expenses to complete the reaction. Consistently with experimental data, we show that the Asp440Ala mutant of the enzyme should exhibit a reduced but retained activity. All considered reaction pathways include proton wires from the O^3'H^3' group via shuttling water molecules.