SNAP-25 S-guanylation and SNARE complex formation

Yusuke Kishimoto, Takaaki Akaike, Hideshi Ihara

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review


8-Nitroguanosine 3′,5′-cyclic monophosphate (8-nitro-cGMP), which is the second messenger in nitric oxide/reactive oxygen species redox signaling, covalently binds to protein thiol groups (called S-guanylation) and exerts various biological functions. Synaptosomal associated protein 25 (SNAP-25), a member of soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins, plays an important role in the process of membrane fusion. We previously showed that SNAP-25 is S-guanylated at cysteine 90. In addition, we revealed that S-guanylation of SNAP-25 increases SNARE complex formation, but decreases the affinity of SNARE complex for complexin. Since SNAP-25 plays a critical role in regulating exocytosis, it is important to elucidate the physiological or pathophysiological meanings of S-guanylation of this protein. Here we describe a protocol for detecting 8-nitro-cGMP and S-guanylated proteins in cells by immunocytochemistry, and methods to detect SNARE complex in 8-nitro-cGMP-treated cells.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Number of pages11
Publication statusPublished - 2019

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745


  • 8-Nitro-cGMP
  • Nitric oxide
  • Redox signal
  • SNAP-25
  • SNARE complex


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