The effect of peroxynitrite on the catalytic activity of soluble guanylyl cyclase.
Soluble guanylyl cyclase (sGC) is a key enzyme of the *NO/cGMP pathway. Many cardiovascular disorders are associated with reduced *NO-mediated effects, while vascular superoxide (O(2)*(-)) production is increased. Both radicals rapidly react to peroxynitrite. We investigated whether peroxynitrite affects the activity and protein expression of sGC in intact vascular preparations. Catalytic sGC activity and expression of the sGC-beta(1) subunit was measured by conversion of radiolabeled GTP and western blot, respectively, using cytosolic extracts from rat aorta that had been incubated for 4 h with *NO/O(2)*(-) systems (devoid of free *NO) generating either 0.13 microM or 7.5 microM peroxynitrite/min. Incubation of rat aorta with 0.13 microM peroxynitrite/min had no effect. In striking contrast, incubation with 7.5 microM peroxynitrite/min resulted in a shift of the concentration-response curve obtained with a *NO donor (p =.0004) and a reduction of maximal specific activity from 3579 +/- 495 to 2422 +/- 265 pmol cGMP/mg/min (p =.036). The expression of the sGC-beta(1) subunit was unchanged. Exposure of aorta to the O(2)*(-) component had no effect, while exposure to the *NO-component reduced sGC expression to 58.8 +/- 7% (p <.001) and maximal sGC activity from 4041 +/- 992 to 1429 +/- 491 pmol cGMP/mg/min (p =.031). These data suggest that continuous generation of extracellular peroxynitrite might interfere with the *NO/cGMP signaling in vascular cells.
Weber
M
M
Institut für Pharmakologie und Klinische Pharmakologie, Medizinische Einrichtungen, Heinrich-Heine-Universität, Düsseldorf, Germany.
Lauer
N
N
Mülsch
A
A
Kojda
G
G
D000818:Animals; D001011:Aorta / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure; D015153:Blotting, Western; D002384:Catalysis; D003600:Cytosol / Q000201:enzymology; D006160:Guanosine Triphosphate / Q000378:metabolism; D006162:Guanylate Cyclase; D008297:Male; D009589:Nitrogen Oxides / Q000378:metabolism; D030421:Peroxynitrous Acid / Q000378:metabolism / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D000071756:Soluble Guanylyl Cyclase; D013481:Superoxides / Q000378:metabolism / Q000494:pharmacology
D009589:Nitrogen Oxides; D018160:Receptors, Cytoplasmic and Nuclear; D013481:Superoxides; D030421:Peroxynitrous Acid; D006160:Guanosine Triphosphate; D006162:Guanylate Cyclase; D000071756:Soluble Guanylyl Cyclase; C039900:nitroxyl