Soluble guanylyl cyclase is a heterodimer of alpha and beta subunits, associated with a prosthetic heme that serves as a receptor for nitric oxide. Since all of these functions require the formation of heterodimers, subunit interactions are an integral part of this analysis.
Nitric oxide is synthesized in most cell types, each responding to diverse stimuli.
Once it is synthesized, NO can act on many different cell types, resulting in a number of possible responses. To examine functions of soluble guanylyl cyclase within one cell type, a dominant negative subunit will be expressed in transgenic mice, using a cell-specific promoter. For example, by cross-breeding these mice with strains that are deficient in parallel redundant signaling mechanisms, we can examine the effect of each pathway alone or in combination during normal and diseased states, such as atherosclerosis. The cortical collecting duct in kidney contains a new form of beta subunit, and we are trying to identify the corresponding alpha subunit in order to study their potential role in hypertension. We are developing a new method to maximize the effectiveness of degenerate primers that are used in the polymerase chain reaction.