Cell-free assay for ubiquitin-independent proteasomal protein degradation.
The ATP-dependent degradation of ornithine decarboxylase is an exceptional case whereby a protein is targeted to the 26S proteasome independently of ubiquitin conjugation. Rather, prior association with the polyamine-induced regulatory protein, antizyme, confers susceptibility of ornithine decarboxylase to proteasomal degradation. In this chapter we describe ornithine decarboxylase/antizyme-based in vivo and in vitro systems for the measurement of ATP-dependent, ubiquitin-independent proteasomal degradation, as well as the application of ornithine decarboxylase as a reporter for the targeting of proteins to the 26S proteasome.
Kahana
Chaim
C
Department of Molecular Genetics, The Weizmann Institute of Science, Rehovot, Israel.
Reiss
Yuval
Y
D000818:Animals; D002460:Cell Line; D002474:Cell-Free System / Q000201:enzymology*; D009955:Ornithine Decarboxylase / Q000737:chemistry*; D046988:Proteasome Endopeptidase Complex; D011817:Rabbits; D012156:Reticulocytes / Q000201:enzymology*; D025801:Ubiquitin / Q000032:analysis* / Q000737:chemistry*
D025801:Ubiquitin; D046988:Proteasome Endopeptidase Complex; D009955:Ornithine Decarboxylase