UNIT 14.5 Analysis of Protein Ubiquitination
Published Online: 1 NOV 2011
Copyright © 2011 by John Wiley & Sons, Inc.
Lab Protocol Title
Current Protocols in Protein Science
How to Cite
Laney, J. D. and Hochstrasser, M. 2011. Analysis of Protein Ubiquitination. Current Protocols in Protein Science. 66:14.5:14.5.1–14.5.13.
- Published Online: 1 NOV 2011
- Published Print: NOV 2011
Attachment of ubiquitin (Ub) to a protein requires a series of enzymes that recognize the substrate and promote Ub transfer. Several methods are described in this unit for determining if a protein has Ub-transferring activity. They include immunoblotting of immunoprecipitated proteins, affinity purification using His-tagged Ub, assaying for auto-ubiquitination of E3, and assaying ubiquitination of a model substrate protein in vitro and in E. coli cells that express Ub-ligation enzymes. These methods are suitable for a variety of eukaryotic cells, but techniques are specifically described for use with yeast and mammalian cells. Curr. Protoc. Protein Sci. 66:14.5.1-14.5.13. © 2011 by John Wiley & Sons, Inc.
- ubiquitin-protein ligase