Assessing Ubiquitylation of Individual Proteins Using Xenopus Extract Systems.
Accepted version
Peer-reviewed
Repository URI
Repository DOI
Change log
Authors
McDowell, Gary S https://orcid.org/0000-0002-9470-3799
Philpott, Anna
Abstract
Xenopus extract systems have been used to study ubiquitylation of proteins, and to uncover some of the fundamental processes of the ubiquitylation pathway itself. They provide a simple, quick, and robust method for studying ubiquitylation. In this protocol, methods are provided for studying protein ubiquitylation using Xenopus egg or embryo extracts and in vitro radiolabeled proteins. These methods also enable examination of whether proteins undergo noncanonical ubiquitylation, through modification of the protein by covalent linkage to ubiquitin through residues other than lysine, such as cysteine, serine, and threonine.
Description
Keywords
Animals, Biochemistry, Biological Assay, Cell Extracts, Protein Biosynthesis, Radiopharmaceuticals, Transcription, Genetic, Ubiquitination, Xenopus Proteins, Xenopus laevis
Journal Title
Cold Spring Harb Protoc
Conference Name
Journal ISSN
1940-3402
1559-6095
1559-6095
Volume Title
2019
Publisher
Cold Spring Harbor Laboratory
Publisher DOI
Sponsorship
Medical Research Council (MC_PC_12009)
Medical Research Council (MR/K018329/1)
Medical Research Council (MR/L021129/1)
Medical Research Council (MR/K018329/1)
Medical Research Council (MR/L021129/1)