Nitric oxide (NO) plays numerous, disparate biological roles that include vasodilation in the cardiovascular system and host defense against microbial pathogens. Nonetheless, the discrete molecular mechanisms involved in NO signaling are not well understood: its molecular relatives S-nitrosothiols (RSNOs) and nitrite (NO2-) can also serve as reservoirs of NO-like behavior. Thus, an understanding of the discrete mechanistic pathways by which these species form, interconvert, and react with molecular targets of biological relevance is crucial to connect nitric oxide to physiological response. Through the use of synthetic models examined in our lab, we share new insights into the interconversion and biological reactivity of key molecules involved in nitric oxide signaling.