@inbook{fd44658448e641d5aad53559d286d967,
title = "SNAP-25 S-guanylation and SNARE complex formation",
abstract = "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.",
keywords = "8-Nitro-cGMP, Nitric oxide, Redox signal, SNAP-25, SNARE complex",
author = "Yusuke Kishimoto and Takaaki Akaike and Hideshi Ihara",
note = "Publisher Copyright: {\textcopyright} Springer Science+Business Media, LLC, part of Springer Nature 2019.",
year = "2019",
doi = "10.1007/978-1-4939-8760-3_9",
language = "English",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "163--173",
booktitle = "Methods in Molecular Biology",
}