title: Effect of S-nitrosothiols on cellular glutathione and reactive protein sulfhydryls.
abstract: S-Nitrosothiols may cause many of the biological effects of NO and cellular effects have been attributed to S-nitrosylation of reactive protein sulfhydryls. This report examines the effect of S-nitrosothiols on the low-molecular-weight thiols and protein thiols in NIH/3T3 cells. A low concentration of S-nitrosocysteine increased the cysteine content of the cells, with no evidence of either low-molecular-weight thiol or protein S-nitrosylation. Millimolar amounts of S-nitrosocysteine produced S-nitrosoglutathione (GSNO), cysteinyl glutathione, cysteine, and glutathione disulfide. Large amounts of protein S-nitrosylation and lesser amounts of protein S-glutathiolation and S-cysteylation were also observed. GSNO and S-nitroso-N-acetylpenicillamine (SNAP) were much less effective than S-nitrosocysteine, but a combination of cysteine and GSNO produced S-nitrosocysteine-like effects. In cultured hepatocytes, millimolar S-nitrosocysteine was significantly less effective since the cells contained three times more glutathione than NIH/3T3 cells. Results suggest that S-nitrosocysteine enters cells intact, and low concentrations do not significantly increase cellular pools of S-nitrosothiol or S-nitrosylated protein. Millimolar concentrations of S-nitrosocysteine generate S-nitrosylated, S-glutathiolated, and S-cysteylated proteins, as well as a variety of low-molecular-weight disulfides and S-nitrosothiols.
authors.affiliation: Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames 50011, USA.
authors.forename: R J
authors.initials: RJ
authors.lastname: Mallis
authors.forename: J A
authors.initials: JA
authors.lastname: Thomas
mesh_terms: D016475:3T3 Cells; D000818:Animals; D005978:Glutathione / Q000378:metabolism*; D051379:Mice; D009603:Nitroso Compounds / Q000494:pharmacology*; D011506:Proteins / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000378:metabolism*
chemical_list: D009603:Nitroso Compounds; D011506:Proteins; D013438:Sulfhydryl Compounds; D005978:Glutathione