We solved the near-atomic resolution structure of smooth muscle myosin-2 in the autoinhibited state (10) using single-particle cryo–electron microscopy. The 3.4-Å structure reveals the precise molecular architecture of 10 and the structural basis for myosin-2 regulation. We reveal the position of the phosphorylation sites that control myosin autoinhibition and activation by phosphorylation of the regulatory light chain. Further, we present a previously unidentified conformational state in myosin-2 that traps ADP and P produced by the hydrolysis of ATP in the active site. This noncanonical state represents a branch of the myosin enzyme cycle and explains the autoinhibition of the enzyme function of 10 along with its reduced affinity for actin. Together, our structure defines the molecular mechanisms that drive 10 formation, stabilization, and relief by phosphorylation of the regulatory light chain.