5
views
0
recommends
+1 Recommend
0 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: not found

      Argininosuccinate synthetase is reversibly inactivated by S-nitrosylation in vitro and in vivo.

      The Journal of Biological Chemistry
      Animals, Aorta, cytology, pathology, Arginine, chemistry, Argininosuccinate Synthase, antagonists & inhibitors, physiology, Blotting, Western, Buthionine Sulfoximine, Catalysis, Cells, Cultured, Cysteine, Dose-Response Relationship, Drug, Glutathione, analogs & derivatives, pharmacology, Glutathione Transferase, metabolism, Humans, Hydrogen Peroxide, Kinetics, Lipopolysaccharides, Male, Mass Spectrometry, Mice, Mice, Inbred C57BL, Muscle, Smooth, Mutagenesis, Site-Directed, Myocytes, Smooth Muscle, Myoglobin, Nitric Oxide, Nitric Oxide Donors, Nitric Oxide Synthase, Nitrogen, Protein Processing, Post-Translational, Rats, Recombinant Proteins, Spectrometry, Mass, Electrospray Ionization, Time Factors

      Read this article at

      ScienceOpenPublisherPubMed
      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          Prior studies have demonstrated that the substrate for NO synthesis, l-arginine, can be regenerated from the NOS co-product l-citrulline. This requires the sequential action of two enzymes, argininosuccinate synthetase (AS) and argininosuccinate lyase (AL). AS activity has been shown to be rate-limiting for high output NO synthesis by immunostimulant-activated cells and represents a potential site for metabolic control of NO synthesis. We now demonstrate that NO mediates reversible S-nitrosylation and inactivation of AS in vitro and in lipopolysaccharide-treated cells and mice. Using a novel mass spectrometry-based method, we show that Cys-132 in human AS is the sole target for S-nitrosylation among five Cys residues. Mutagenesis studies confirm that S-nitrosylation of Cys-132 is both necessary and sufficient for the inhibition of AS by NO donors. S-nitroso-AS content is regulated by cellular glutathione levels and selectively influences NO production when citrulline is provided to cells as a protosubstrate of NOS but not when l-arginine is provided. A phylogenetic comparison of AS sequences suggests that Cys-132 evolved as a site for post-translational regulation of activity in the AS in NOS-expressing species, endowing NO with the capacity to limit its own synthesis by restricting arginine availability.

          Related collections

          Author and article information

          Comments

          Comment on this article