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Detection and Analysis of Protein ISGylation
Abstract
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.
Affiliation(s): (3) Department of Biochemistry, Graduate School of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan
Series: Methods in Molecular Biology  |  Volume: 446  |  Pub. Date: Apr-04-2008  |  Page Range: 139-149  |  DOI: 10.1007/978-1-60327-084-7_10
Subject:  Protein Science
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