Guanylyl cyclases (GC) are proteins that are essential for the production of the second messenger cyclic guanosine monophosphate (cGMP). Mammalian GC have attracted considerable interest due to their roles in important physiological processes such as vasodilation, vision, and bone growth. In addition, their link to disease and concomitant pharmaceutical potential have made these cyclases a long standing target for probing their intriguing mechanism of activation with the aim of drug development. The vasodilatory drugs nitroglycerin and nesiritide act through (different) GC pathways and have both been shown to provide beneficial relief for congestive heart failure patients. New structural insights are recently emerging on the activation mechanism and regulation of these receptors. The aim of this review is to discuss the interesting differences and similarities between members of the soluble and membrane bound GC in detail and put these in context with the structural knowledge that is available to date. These efforts contribute to an enhanced understanding of the GC and will likely lead to an increased success in structure-based therapeutic intervention.