Detection of modification by ubiquitin-like proteins.
Ubiquitin and ubiquitin-like proteins (Ubls) are conjugated to many target proteins either as monomeric units or as polymeric chains. There are at least 12 members of the ubiquitin family in the human genome and their conjugation dramatically alters the properties of the modified protein. The presence of highly active proteases that specifically deconjugate Ubls often means that, in the cell, the steady state level of modified protein is low. Detection of protein species modified by Ubls can therefore represent a significant challenge. Here, we describe methods that have been developed to allow detection of Ubl modified proteins both in vivo and in vitro.
Jaffray
Ellis G
EG
Centre for Biomolecular Science, School of Biology, University of St. Andrews, North Haugh, St. Andrews, KY16 9ST, UK.
Hay
Ronald T
RT
D015153:Blotting, Western; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D006639:Histidine; D006801:Humans; D047468:Immunoprecipitation; D008954:Models, Biological; D009842:Oligopeptides; D011480:Protease Inhibitors / Q000494:pharmacology; D011489:Protein Denaturation; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000032:analysis / Q000378:metabolism; D014452:Ubiquitins / Q000032:analysis / Q000378:metabolism*
C471213:His-His-His-His-His-His; D009842:Oligopeptides; D011480:Protease Inhibitors; D011506:Proteins; D014452:Ubiquitins; D006639:Histidine