Abstract
Interferon-stimulated gene 15 (ISG15) encodes a protein that is most upregulated by type I interferon stimulation and, upon activation, is conjugated to various target proteins in a process known as ISGylation. ISGylation has been shown to have roles in various biological phenomena such as viral infection and cancer. To gain further insight into the function of ISGylation, it would be useful to be able to identify ISGylated proteins. Here, we describe a method for the identification of proteins modified with ISG15. This method involves the generation of stable ISG15-transfectant cells, followed by affinity purification, and then identification of the ISGylated proteins by mass spectrometry.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Zhang D, Zhang DE (2011) Interferon-stimulated gene 15 and the protein ISGylation system. J Interf Cytokine Res 31(1):119–130
Knight E Jr, Fahey D, Cordova B et al (1988) A 15-kDa interferon-induced protein is derived by COOH-terminal processing of a 17-kDa precursor. J Biol Chem 263(10):4520–4522
Narasimhan J, Potter JL, Haas AL (1996) Conjugation of the 15-kDa interferon-induced ubiquitin homolog is distinct from that of ubiquitin. J Biol Chem 271(1):324–330
Malakhova O, Malakhov M, Hetherington C et al (2002) Lipopolysaccharide activates the expression of ISG15-specific protease UBP43 via interferon regulatory factor 3. J Biol Chem 277(17):14703–14711
Zhao C, Collins MN, Hsiang TY et al (2013) Interferon-induced ISG15 pathway: an ongoing virus-host battle. Trends Microbiol 21(4):181–186
Sgorbissa A, Brancolini C (2012) IFNs, ISGylation and cancer: Cui prodest? Cytokine Growth Factor Rev 23(6):307–314
Bogunovic D, Byun M, Durfee LA et al (2012) Mycobacterial disease and impaired IFN-gamma immunity in humans with inherited ISG15 deficiency. Science 337(6102):1684–1688
Zhao C, Denison C, Huibregtse JM et al (2005) Human ISG15 conjugation targets both IFN-induced and constitutively expressed proteins functioning in diverse cellular pathways. Proc Natl Acad Sci U S A 102(29):10200–10205
Giannakopoulos NV, Luo JK, Papov V et al (2005) Proteomic identification of proteins conjugated to ISG15 in mouse and human cells. Biochem Biophys Res Commun 336(2):496–506
Takeuchi T, Inoue S, Yokosawa H (2006) Identification and Herc5-mediated ISGylation of novel target proteins. Biochem Biophys Res Commun 348(2):473–477
Takeuchi T, Yokosawa H (2005) ISG15 modification of Ubc13 suppresses its ubiquitin-conjugating activity. Biochem Biophys Res Commun 336(1):9–13
Ohtsuka T, Nakanishi H, Ikeda W et al (1998) Nexilin: a novel actin filament-binding protein localized at cell-matrix adherens junction. J Cell Biol 143(5):1227–1238
Wang W, Zhang W, Han Y et al (2005) NELIN, a new F-actin associated protein, stimulates HeLa cell migration and adhesion. Biochem Biophys Res Commun 330(4):1127–1131
Desai SD, Reed RE, Burks J et al (2012) ISG15 disrupts cytoskeletal architecture and promotes motility in human breast cancer cells. Exp Biol Med (Maywood) 237(1):38–49
Thiede B, Hohenwarter W, Krah A (2005) Peptide mass fingerprinting. Methods 35(3):237–247
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Takeuchi, T., Koinuma, S., Yokosawa, H., Arata, Y. (2019). An Approach for the Identification of Proteins Modified with ISG15. In: Kannicht, C. (eds) Post-Translational Modification of Proteins. Methods in Molecular Biology, vol 1934. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9055-9_15
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9055-9_15
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9053-5
Online ISBN: 978-1-4939-9055-9
eBook Packages: Springer Protocols