title: Basal and stimulated protein S-nitrosylation in multiple cell types and tissues.
abstract: There is substantial evidence that protein S-nitrosylation provides a significant route through which nitric oxide (NO)-derived bioactivity is conveyed. However, most examples of S-nitrosylation have been characterized on the basis of analysis in vitro, and relatively little progress has been made in assessing the participant forms of nitric-oxide synthase (NOS) or the dynamics of protein S-nitrosylation in situ. Here we utilize antibodies specific for the nitrosothiol (SNO) moiety to provide an immunohistochemical demonstration that protein S-nitrosylation is coupled to the activity of each of the major forms of NOS. In cultured endothelial cells, SNO-protein immunoreactivity increases in response to Ca(2+)-stimulated endothelial NOS (eNOS) activity, and in aortic rings, endothelium-derived and eNOS-mediated relaxation (EDRF) is coupled to increased protein S-nitrosylation in both endothelial and associated smooth muscle cells. In cultured macrophages, SNO-protein levels increase upon cytokine induction of induced NOS (iNOS), and in PC12 cells, increased protein S-nitrosylation is linked to nerve growth factor induction of neuronal NOS (nNOS). In addition, we describe developmental and pathophysiological increases in SNO-protein immunoreactivity within human lung. These results, which demonstrate Ca(2+), neurohumoral, growth factor, cytokine, and developmental regulation of protein S-nitrosylation that is coupled to NOS expression and activity, provide unique evidence for the proposition that this ubiquitous NO-derived post-translational protein modification serves as a major effector of NO-related bioactivity.
authors.affiliation: Department of Medicine and Howard Hughes Medical Institute, Duke University Medical Center, Durham, North Carolina 27710, USA.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Gow
authors.forename: Qiping
authors.initials: Q
authors.lastname: Chen
authors.forename: Douglas T
authors.initials: DT
authors.lastname: Hess
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Day
authors.forename: Harry
authors.initials: H
authors.lastname: Ischiropoulos
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Stamler
mesh_terms: D000818:Animals; D001011:Aorta / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D002460:Cell Line; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005227:Fatty Acids / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D007525:Isoelectric Focusing; D008168:Lung / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism; D051379:Mice; D017942:Nitric Acid / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D016716:PC12 Cells; D011485:Protein Binding; D051381:Rats; D026422:S-Nitrosoglutathione / Q000378:metabolism; D013552:Swine
chemical_list: D005227:Fatty Acids; D017942:Nitric Acid; D026422:S-Nitrosoglutathione; D019001:Nitric Oxide Synthase; D002118:Calcium