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 alpha1/beta1 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.
Martin
Emil
E
Department of Integrative Biology and Pharmacology, UT Health Science Center at Houston Medical School, Houston, TX 77030, USA.
Berka
Vladimir
V
Tsai
Ah-Lim
AL
Murad
Ferid
F
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D002248:Carbon Monoxide / Q000378:metabolism; D002460:Cell Line; D017931:DNA Primers; D004578:Electron Spin Resonance Spectroscopy; D006162:Guanylate Cyclase; D006418:Heme / Q000096:biosynthesis / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D009569:Nitric Oxide / Q000378:metabolism*; D011485:Protein Binding; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D000071756:Soluble Guanylyl Cyclase; D018411:Spodoptera
D017931:DNA Primers; D018160:Receptors, Cytoplasmic and Nuclear; D011994:Recombinant Proteins; D009569:Nitric Oxide; D006418:Heme; D002248:Carbon Monoxide; D006162:Guanylate Cyclase; D000071756:Soluble Guanylyl Cyclase