Summary
ISG15 is a ubiquitin-like modifi er that is conjugated to target proteins by a sequential reaction catalyzed by E1/E2/E3 enzymes (protein ISGylation). ISG15 and protein ISGylation are upregulated by interferon stimuli. ISG15 functions as an antiviral protein against Sindbis virus and HIV-1, but the molecular mechanism remains unknown. Here we describe in detail methods for detecting and analyzing protein ISGylation. The methods consist of plasmid transfection and affi nity purifi cation of ISGylated proteins. In addition, we describe a method for detecting ISGylation of a target protein, Ubc13.
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Acknowledgments
This study was supported in part by grants-in-aid for scientific research from the Ministry of Education, Culture, Sports, Science and Technology of Japan.
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© 2008 Humana Press, a part of Springer Science+Business Media, LLC
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Takeuchi, T., Yokosawa, H. (2008). Detection and Analysis of Protein ISGylation. In: Kannicht, C. (eds) Post-translational Modifications of Proteins. Methods in Molecular Biology™, vol 446. Humana Press. https://doi.org/10.1007/978-1-60327-084-7_10
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DOI: https://doi.org/10.1007/978-1-60327-084-7_10
Publisher Name: Humana Press
Print ISBN: 978-1-58829-719-8
Online ISBN: 978-1-60327-084-7
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