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Assaying Proteasomal Degradation in a Cell-free System in Plants
Published on: March 26, 2014
Cell-free assay for ubiquitin-independent proteasomal protein degradation
1Department of Molecular Genetics, The Weizmann Institute of Science, Rehovot, Israel.
Abstract:
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.
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