Abstract
S-nitrosylation, which involves the coupling of an NO group to the reactive thiol of Cys residue(s) in a polypeptide, is an important posttranslational modification detected in a variety of proteins. Here, we present the S-nitrosylation of recombinant galectin-2 (Gal-2) using S-nitrosocysteine and the measurement of the molecular ratio of S-nitrosylation of Cys residues in the Gal-2 protein.
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References
Oka T, Murakami S, Arata Y et al (1999) Identification and cloning of rat galectin-2: expression is predominantly in epithelial cells of the stomach. Arch Biochem Biophys 361:195–201
Nio-Kobayashi J, Takahashi-Iwanaga H, Iwanaga T (2009) Immunohistochemical localization of six galectin subtypes in the mouse digestive tract. J Histochem Cytochem 57:41–50
Saal I, Lensch M, Lohr M et al (2005) Human galectin-2: expression profiling by RT-PCR/immunohistochemistry and its introduction as a histochemical tool for ligand localization. Histol Histopathol 20:1191–1208
Ohtake K, Shimada N, Uchida H et al (2009) Proteomic approach for identification of protein S-nitrosation in mouse gastric mucosa treated with S-nitrosoglutathione. J Proteome 72:750–760
Hess DT, Matsumoto A, Kim SO et al (2005) Protein S-nitrosylation: purview and parameters. Nat Rev Mol Cell Biol 6:150–166
Tamura M, Saito M, Yamamoto K et al (2015) S-nitrosylation of mouse galectin-2 prevents oxidative inactivation by hydrogen peroxide. Biochem Biophys Res Commun 457:712–717
Tamura M, Sasai A, Ozawa R et al (2016) Identification of the cysteine residue responsible for oxidative inactivation of mouse galectin-2. J Biochem 160:233–241
Sakakura M, Tamura M, Fujii N et al (2018) Structural mechanisms for the S-nitrosylation-derived protection of mouse galectin-2 from oxidation-induced inactivation revealed by NMR. FEBS J 285:1129–1145
Saville BB (1958) A scheme for the colorimetric determination of microgram amounts of thiols. Analyst 83:670–672
Griess P (1879) Amidosäuren mit Alkoholradikalen. Chem Ber 12:426–428
Williams DL (1996) S-Nitrosothiols and role of metal ions in decomposition to nitric oxide. Methods Enzymol 268:299–308
Hoffmann J, Dimmeler S, Haendeler J (2003) Shear stress increases the amount of S-nitrosylated molecules in endothelial cells: important role for signal transduction. FEBS Lett 551:153–158
Gordge MP, Meyer DJ, Hothersall J et al (1995) Copper chelation-induced reduction of the biological activity of S-nitrosothiols. Br J Pharmacol 114:1083–1089
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Tamura, M., Arata, Y. (2020). Expression, S-Nitrosylation, and Measurement of S-Nitrosylation Ratio of Recombinant Galectin-2. In: Hirabayashi, J. (eds) Lectin Purification and Analysis. Methods in Molecular Biology, vol 2132. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0430-4_6
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DOI: https://doi.org/10.1007/978-1-0716-0430-4_6
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0429-8
Online ISBN: 978-1-0716-0430-4
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