Interaction between nitric oxide and prostaglandin H synthase.
Prostaglandin H synthase (PGHS) is a hemeprotein, and thus its catalytic activity potentially could be modulated by direct interaction with nitric oxide (NO). We have monitored spectroscopic and activity changes in pure ovine PGHS isoform-1 to investigate its interaction with NO in more detail. The binding kinetics for NO and the ferric heme in resting PGHS were analyzed by stopped-flow spectrophotometry at 21 degrees C. The rate constants for association and dissociation were estimated to be 6.5 x 10(4) M-1 s-1 and 60 s-1, respectively, leading to an equilibrium dissociation constant (Kd) of 0.92 mM. NO thus has a relatively weak affinity for heme in ferric PGHS, the resting oxidation state of this hemeprotein. NO did react strongly and completely with ferrous PGHS under anaerobic conditions, displacing the proximal histidine ligand to the prosthetic group. Dissolved NO at up to 2 mM produced only slight decreases in the cyclooxygenase activity of microsomal, detergent-extracted, or homogeneous preparations of ovine PGHS. The NO donors sodium nitroprusside and glyceryl trinitrate at levels of up to 1 mM also had little effect on the activity of the PGHS preparations. Thus, there was no evidence for significant direct interaction of PGHS with NO at concentrations likely to be encountered in vivo.
Tsai
A L
AL
Department of Internal Medicine, University of Texas Health Science Center at Houston 77030.
Wei
C
C
Kulmacz
R J
RJ
D000818:Animals; D004578:Electron Spin Resonance Spectroscopy; D006418:Heme / Q000378:metabolism; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008297:Male; D009569:Nitric Oxide / Q000378:metabolism* / Q000494:pharmacology; D005996:Nitroglycerin / Q000494:pharmacology; D009599:Nitroprusside / Q000494:pharmacology; D011451:Prostaglandin-Endoperoxide Synthases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D012669:Seminal Vesicles / Q000201:enzymology; D012756:Sheep; D013053:Spectrophotometry
D007527:Isoenzymes; D009599:Nitroprusside; D009569:Nitric Oxide; D006418:Heme; D011451:Prostaglandin-Endoperoxide Synthases; D005996:Nitroglycerin