Analysis of protein ubiquitination.

Jeffrey D Laney, Mark Hochstrasser

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Attachment of ubiquitin (Ub) to a protein requires a complex of enzymes that recognize the substrate and promote Ub transfer. Sequence motifs present in these enzymes may indicate that other uncharacterized proteins containing these motifs have a biochemical function of Ub-protein ligation, and several in vitro methods are described in this unit for determining if a protein has Ub-transferring activity. They include psmunoblotting of psmunoprecipitated proteins, affinity purification using His-tagged ubiquitin, assaying for auto-ubiquitination of E3, and assaying ubiquitination of a model substrate protein. These methods are suitable for a variety of eukaryotic cells, but techniques are specifically described for use with yeast and mammalian cells.

Original languageEnglish (US)
JournalCurrent Protocols in Protein Science
VolumeChapter 14
StatePublished - Nov 2002
Externally publishedYes

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Ubiquitination
Ubiquitin
Proteins
Amino Acid Motifs
Eukaryotic Cells
Enzymes
Ligation
Substrates
Yeast
Yeasts
Purification
Cells

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Analysis of protein ubiquitination. / Laney, Jeffrey D; Hochstrasser, Mark.

In: Current Protocols in Protein Science, Vol. Chapter 14, 11.2002.

Research output: Contribution to journalArticle

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