Article thumbnail

S-Nitrosation of Cellular Proteins by NO Donors in Rat Embryonic Fibroblast 3Y1 Cells: Factors Affecting S-Nitrosation

By Norihiro Ryuman, Nobuo Watanabe and Takao Arai


The mechanism of protein S-nitrosation in cells is not fully understood. Using rat 3Y1 cells, we addressed this issue. Among S-nitrosothiols and NO donors tested, only S-nitrosocysteine (CysNO) induced S-nitrosation when exposed in Hanks' balanced salt solution (HBSS) and not in serum-containing general culture medium. In HBSS, NO release from CysNO was almost completely abolished by sequestering metal ions with a metal chelator without affecting cellular S-nitrosation. In contrast, L-leucine, a substrate of L-type amino acid transporters (LATs), significantly inhibited S-nitrosation. The absence of S-nitrosation with CysNO in general culture medium resulted not only from a competition with amino acids in the medium for LATs but also from transnitrosation of cysteine residues in serum albumin. Collectively, these results suggest that in simple buffered saline, CysNO-dependent S-nitrosation occurs through a cellular incorporation-dependent mechanism, but if it occurs in general culture media, it may be through an NO-dependent mechanism

Topics: Research Article
Publisher: Hindawi Publishing Corporation
OAI identifier:
Provided by: PubMed Central

To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.

Suggested articles


  1. (2001). a m a c h a n d r a n ,P .R o o t ,X .M .J i a n g ,P .J .H o g g ,a n dB . Mutus, “Mechanism of transfer of NO from extracellular Snitrosothiols into the cytosol by cell-surface protein disulfide isomerase,”
  2. (1995). A new nitric oxide (NO) releaser: spontaneous
  3. (2009). Activation and inhibition of soluble guanylyl cyclase by Snitrosocysteine: involvement of amino acid transport system L,” Free Radical Biology and Medicine,
  4. (2009). B r y a n ,K .B i a n ,a n dF .M u r a d ,“ D i s c o v e r yo ft h en i t r i c oxide signaling pathway and targets for drug development,”
  5. (1996). C o o k ,S .Y .K i m ,D .T e a g u ee ta l . ,“ C o n v e n i e n tc o l o r i -metric and fluorometric assays for S-nitrosothiols,”
  6. (2002). Catalysis of S-nitrosothiols formation by serum albumin: the mechanism and implication in vascular control,”
  7. (1999). Cellsurface protein disulfide isomerase catalyzes transnitrosation and regulates intracellular transfer of nitric oxide,”
  8. cytotoxicity to PC12cellsismediatedbythiol-sensitiveshort-livedsubstances generated through SIN-1 decomposition in culture medium,”
  9. (2009). Dinitrosyliron complexes and the mechanism(s) of cellular protein nitrosothiol formation from nitric
  10. (2009). e l v a k u m a r ,R .L .H u g a n i r ,a n dS .H .S n y d e r ,“ S - n i t r o -sylation of stargazin regulates surface expression of AMPAglutamate neurotransmitter receptors,”
  11. (1996). e s h i v e ,S .S i n g h ,J .S .W i s h n o k ,S .R .T a n n e n b a u m ,a n d
  12. (2009). Effects of uric acid on nitrosation of N-acetylcysteine by diethylamine nonoate and N-acetyl-N-nitrosotryptophan,”
  13. (2006). Extracellular S-nitrosoglutathione, but not S-nitrosocysteine or N 2O3, mediates protein S-nitrosation in rat spinal cord slices,”
  14. (1998). Fluorometric detection of
  15. (2007). Functional characterization of two S-nitroso-L-cysteine transporters, which mediate movement of NO equivalents into vascular cells,”
  16. (2008). h u ,S .L i ,Z .M .M a r s h a l l ,a n dA .R .W h o r t o n ,“ Ac y s t i n e -cysteine shuttle mediated by xCT facilitates cellular responses to S-nitrosoalbumin,”
  17. (1975). i m u r a ,A .I t a g a k i ,a n dJ .S u m m e r s ,“ R a tc e l ll i n e3 Y 1 and its virogenic polyoma and SV50 transformed derivatives,”
  18. (2009). Identification of major Snitrosylated proteins in murine experimental autoimmune encephalomyelitis,”
  19. (2005). Identification of stereoselective transporters for S-nitroso-L-cysteine: role of LAT1 and LAT2 in biological activity of S-nitrosothiols,”
  20. (2007). Interactions between cell surface protein disulphide isomerase and S-nitrosoglutathione during nitric oxide delivery,”
  21. (1997). Involvement of L-typelike amino acid transporters in S-nitrosocysteine-stimulated noradrenaline release in the rat hippocampus,”
  22. (2007). J.Mannick,andB.Gaston,“S-Nitrosothiol measurements in biological systems,”
  23. (2007). M¨ o l l e r ,Q .L i ,J .R .L a n c a s t e rJ r . ,a n dA .D e n i c o l a , “Acceleration of nitric oxide autoxidation and nitrosation by membranes,”
  24. (2010). Nitrosation of N-methyl derivatives of uric acid and their transnitrosation ability to
  25. (2008). Nitrosothiol reactivity profiling identifies S-nitrosylated proteins with unexpected stability,”
  26. (2001). Protein S-nitrosylation: a physiological signal for neuronal nitric oxide,”
  27. (2008). Pulmonary alveolar epithelial uptake of S-nitrosothiols is regulated by Ltype amino acid transporter,”
  28. (2006). Regulation and specificity of S-nitrosylation and denitrosylation,”
  29. (2006). Requirement of transmembrane transport for S-nitrosocysteine-dependent modification of intracellular thiols,”
  30. (2008). S-nitrosylated human serum albumin-mediated cytoprotective activity is enhanced by fatty acid binding,” J o u r n a lo fB i o l o g i c a lC h e m -istry,
  31. (2009). S-nitrosylation of Drp1 mediates β-amyloid-related mitochondrial fission and neuronal injury,”
  32. (2008). S-nitrosylation of histone deacetylase 2 induces chromatin remodelling in neurons,”
  33. (1991). Signal transduction mechanisms involving nitric oxide,”
  34. (2005). Stamler,“Protein S-nitrosylation:purview and parameters,”
  35. (2002). Stoclet,“Evidence thatintrinsiciron butnotintrinsic copper determines S-nitrosocysteine decomposition in buffer solution,”
  36. (2004). The mechanism of transmembrane S-nitrosothioltransport,”
  37. (2005). Y.YangandJ.Loscalzo,“S-nitrosoproteinformationandlocalization in endothelial cells,”
  38. (2006). Yaoetal.,“S-Nitrosylated proteindisulphide isomerase links protein misfolding to neurodegeneration,”
  39. (1996). Z h a n g ,A .M .X u ,M .N o m e n ,M .W a l s h ,J .F .K e a n e y ,a n d