Identifying a Ubiquitinated Adaptor Protein by a Viral E3 Ligase Through Co-immunoprecipitation

Chenhe Su; Chenhao Su; Chunfu Zheng

doi:10.1007/978-1-0716-4108-8_5

Identifying a Ubiquitinated Adaptor Protein by a Viral E3 Ligase Through Co-immunoprecipitation

Methods Mol Biol. 2025:2854:35-40. doi: 10.1007/978-1-0716-4108-8_5.

Authors

Chenhe Su¹, Chenhao Su², Chunfu Zheng³

Affiliations

¹ State Key Laboratory of Antiviral Drugs, Pingyuan Laboratory, NMPA Key Laboratory for Research and Evaluation of Innovative Drug, School of Chemistry and Chemical Engineering, Henan Normal University, Xinxiang, Henan, China.
² Department of Nephrology and Rheumatology, The Second Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
³ Department of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, AB, Canada.

PMID: 39192116
DOI: 10.1007/978-1-0716-4108-8_5

Abstract

Co-immunoprecipitation is a technique widely utilized to isolate protein complexes and study protein-protein interactions. Ubiquitinated proteins could be identified by combining co-immunoprecipitation with SDS-PAGE followed by immunoblotting. In this chapter, we use Herpes Simplex Virus 1 immediate-early protein ICP0-mediated polyubiquitination of p50 as an example to describe the method to identify a ubiquitinated adaptor protein by a viral E3 ligase by co-immunoprecipitation.

Keywords: A signaling adaptor; Co-immunoprecipitation; Ubiquitination; Viral E3 ligase.

MeSH terms

Electrophoresis, Polyacrylamide Gel / methods
Herpesvirus 1, Human / metabolism
Humans
Immediate-Early Proteins* / metabolism
Immunoprecipitation* / methods
Protein Binding
Ubiquitin-Protein Ligases* / metabolism
Ubiquitinated Proteins / metabolism
Ubiquitination*
Viral Proteins / metabolism

Substances

Ubiquitin-Protein Ligases
Immediate-Early Proteins
Vmw110 protein, Human herpesvirus 1
Ubiquitinated Proteins
Viral Proteins