Format

Send to

Choose Destination
See comment in PubMed Commons below
Methods Enzymol. 2005;398:68-73.

In vitro systems for NEDD8 conjugation by Ubc12.

Author information

1
Laboratory of Frontier Science, The Tokyo Metropolitan Institute of Medical Science, Honkomagome, Bunkyo-ku, Tokyo 113-8613, Japan.

Abstract

Nedd8 is a ubiquitin-like molecule that is highly conserved in eukaryotes. Similar to ubiquitin, Nedd8 attaches to target proteins through an enzymatic cascade composed of Nedd8-specific E1 (activating)- and E2 (conjugating)-enzymes. The E1 for Nedd8 is a heterodimer of APP-BP1 and Uba3, while the E2 is Ubc12. The most well-characterized targets of Nedd8 are proteins of the Cullin family, a core component of SCF (Skp1/Cullin1/F-box proteins) and/or SCF-like ubiquitin ligase complexes. The Nedd8 modification of Cullin (Cul) family proteins is evolutionarily conserved, and genetic analyses in various organisms suggest a positive role of the NEDD8 for the function of Cul family proteins. Further biochemical analysis reveals that NEDD8 modification augments the ubiquitin ligase activity of Cullin-based complexes through the recruitment of ubiquitin-charged E2 to the complex. This chapter describes methods for the purification of NEDD8 conjugation enzymes and in vitro Nedd8 conjugation.

PMID:
16275320
DOI:
10.1016/S0076-6879(05)98007-5
[Indexed for MEDLINE]
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Elsevier Science
    Loading ...
    Support Center