Identification of S-nitrosylated proteins in endotoxin-stimulated RAW264.7 murine macrophages.

Journal Article (Journal Article)

Nitric oxide (NO) is an omnipresent regulator of cell function in a variety of physiologic and pathophysiologic states. In part, NO exerts its actions by S-nitrosylation of target thiols, primarily in cysteine residues. Delineating the functional correlates of S-nitrosylation can begin with identification of the entire population of S-nitrososylated proteins. Recently, the biotin switch technique was developed to allow a proteomic approach to identification of the "universe" of S-nitrsoylated proteins. In this study using endotoxin-stimulated RAW264.7 murine macrophages, we have utilized the biotin-switch technique and protein sequencing to identify S-nitrosylated proteins in this setting. In contrast to other studies utilizing exogenous sources of NO, our approach utilizes endogenous NO synthesis as the basis for S-nitrosylation. Our results indicate multiple unique proteins not previously identified as S-nitrosylation targets: enolase, pyruvate kinase, elongation factor-1 and -2, plastin-2, FRAG-6, CEM-16, and SMC-6. While the ubiquitous nature of NO argues for some degrees of commonality, S-nitrosylation of unique proteins specific to endotoxin stimulated macrophages suggests regulatory mechanisms for which NO is necessary, but not sufficient.

Full Text

Duke Authors

Cited Authors

  • Gao, C; Guo, H; Wei, J; Mi, Z; Wai, PY; Kuo, PC

Published Date

  • March 2005

Published In

Volume / Issue

  • 12 / 2

Start / End Page

  • 121 - 126

PubMed ID

  • 15740986

International Standard Serial Number (ISSN)

  • 1089-8603

Digital Object Identifier (DOI)

  • 10.1016/j.niox.2004.11.006


  • eng

Conference Location

  • United States