From biochemical and structural studies of soluble guanylate cyclase toward drug design

To achieve this goal, I developed a heterologous expression system of full-length bovine sGC (Fl-sGC). Early attempts to produce recombinant bovine sGC in E. coli resulted in misfolded protein accumulation. Indeed, producing soluble protein in Escherichia coli is still a major difficulty in the sGC field. By using fusion technology, I successfully overexpressed both α and β subunits in a soluble heme-bound active form. Optimization of expression levels by varying bacterial growth conditions including temperature, media, additives and induction, will be followed by purification and characterization of Fl-sGC. So, a crucial step has been achieved, allowing us to pursue structural studies to probe the structure and mechanism of sGC and promote the discovery of stimulators of this physiologically important enzyme. from 4th International Conference of cGMP Generators, Effectors and Therapeutic Implications Regensburg, Germany. 19–21 June 2009