Soluble guanylyl cyclase: The nitric oxide receptor

Soluble guanylyl cyclase is recognized as the most sensitive physiologic receptor for nitric oxide. Binding of nitric oxide to the heme moiety of the cyclase induces its capacity to synthesize the second messenger cGMP. Although the changes in the state of the heme moiety upon exposure of enzyme to NO and its correlation to the stimulation of sGC catalytic activity are well documented, the exact mechanism of such coupling is not understood. Structure-functional studies are required to elucidate this process. In this chapter, we describe the method of expression and purification of recombinant human alpha(1)/beta(1) isoform of sGC in insect cells, which can be a useful tool for such studies. Several approaches that enable characterization of the binding of NO to sGC heme moiety are also described.