Guanylyl cyclases (GCs) catalyze the conversion of GTP to the second messenger cGMP. While some transmembrane GCs are receptors for extracellular ligands, other transmembrane GCs such as retinal-specific GC-E and GC-F are stimulated by cellular proteins. GC-D is expressed in a special group of olfactory sensory neurons. However, the direct regulatory mechanism of GC-D activity is not completely understood. Here we have demonstrated that bicarbonate directly increases the activity of purified GC-D. Bicarbonate also increases the cGMP levels in cells expressing GC-D. These results identify bicarbonate as a small molecule that regulates GC-D.