Synergistic Interplay of Stimulatory Cofactors in the Activation of Adenylyl Cyclase Isoform 1

Adenylyl Cyclase isoform 1 (AC1), responsible for synthesizing the signaling molecule cyclic adenosine monophosphate (cAMP), is key in synaptic plasticity, long-term chronic pain syndromes, osteosarcoma-associated pain, and drug abuse. The protein calmodulin (CaM) and the small molecule forskolin (Fsk) stimulate AC1 to form a catalytic site for ATP catalysis; however, molecular AC1 structural changes triggered by CaM and Fsk remain poorly understood. This study developed a computational model for AC1–cofactor complexes and used all-atom molecular dynamics (MD) simulations to determine how CaM and Fsk individually and jointly affect AC1 dynamics and assess whether they exhibit any synergistic effects. Four systems were investigated: AC1-No Partner, AC1-CaM, AC1-Fsk, and AC1-CaM-Fsk. Simulations revealed that individual and joint cofactor bindings induced unique structural changes within the regulatory C1b subdomain of AC1. CaM and Fsk binding result in a reduced cross section of the catalytic site, implying tighter binding for ATP. Notably, the results showed that the AC1-CaM-Fsk system exhibited unique features distinct from the AC1-CaM and AC1-Fsk systems, demonstrating synergistic effects of CaM and Fsk. Our understanding of AC1-cofactor interactions can guide future research toward modulating AC1 activity, potentially contributing to the development of novel treatments for AC1-associated diseases.