Purified nitric oxide synthase

Abstract

The present invention provides a novel constitutive nitric oxide synthase (NOS) that utilizes both L-arginine and arginine-rich peptides, oligopeptides or proteins, e.g., bradykinin (BK), as substrates in the synthesis of nitric oxide (NO). Also provided are methods of controlling, regulating or modulating NO synthesis in a subject.

What is claimed is:  
     
         1 . A method of regulating or controlling nitric oxide production in a mammalian subject comprising administering to the mammal a nitric oxide-regulating amount of an arginine-rich peptide, oligopeptide, or protein inhibitor of nitric oxide synthase.  
     
     
         2 . The method of  claim 1 , wherein the nitric oxide synthase is nNOS-II.  
     
     
         3 . The method of  claim 1 , wherein the inhibitor is N G -methyl-L-arginine.  
     
     
         4 . The method of  claim 1 , wherein the inhibitor is N G -nitro-L-arginine.  
     
     
         5 . The method of  claim 1 , wherein the inhibitor is a peptide or oligopeptide.  
     
     
         6 . The method of  claim 1 , wherein the nitric oxide production is increased.  
     
     
         7 . The method of reducing the rate of nitric oxide production in a mammalian subject comprising administering to the mammal a nitric oxide inhibiting amount of a peptide, oligopeptide, or protein inhibitor of the nitric oxide synthase of  claim 1 .  
     
     
         8 . The method of reducing the rate of nitric oxide production in a mammalian subject comprising administering to the mammal a nitric oxide inhibiting amount of a peptide, oligopeptide, or protein inhibitor of the nitric oxide synthase of  claim 2 .  
     
     
         9 . The method of  claim 1 , wherein the nitric oxide production is decreased.  
     
     
         10 . A method of preventing or treating a nitric oxide-mediated disease or condition in a mammalian subject comprising administering to the subject in need of such prevention or treatment a therapeutically effective amount of a peptide, oligopeptide or protein inhibitor of nitric oxide synthase.  
     
     
         11 . The method of  claim 10 , wherein the nitric oxide synthase is nNOS-II.  
     
     
         12 . The method of  claim 10 , wherein the nitric oxide synthase is a mammalian brain-derived nitric oxide synthase (NOS) protein purified to an activity at least 6,360-fold, said protein having a denatured molecular mass as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis under reducing conditions of about 105 kD, and a native homodimeric molecular mass as determined by gel filtration of about 230 kD), requiring FAD, FMN, Ca 2+  and tetrahydrobiopterin cofactors for the production of nitric oxide either from L-arginine, or an analog or derivative thereof, or from an arginine-rich peptide, oligopeptide, or protein substrate.  
     
     
         13 . The method of  claim 10 , wherein the inhibitor is N G -methyl-L-arginine.  
     
     
         14 . The method of  claim 10 , wherein the inhibitor is N G -nitro-L-arginine.  
     
     
         15 . The method of  claim 10 , wherein the inhibitor is a peptide or oligopeptide.