title: Protocols for studying antizyme expression and function.
abstract: Antizyme (AZ) is a key molecule in feedback regulation of cellular polyamines. It is induced by polyamines through stimulation of ribosomal frameshifting during its translation. In mammals, AZ is diverged into three paralogs, AZ1-3. Tissue and subcellular distribution are different among the paralogs, as determined by immunochemical methods or expression of fluorescent-tagged proteins. Only AZ2 is known to be phosphorylated. AZ regulates cellular polyamine levels through multiple mechanisms. It binds to ornithine decarboxylase (ODC) to form an inactive complex and to trigger degradation of ODC by 26S proteasomes. The AZ activity to promote ODC degradation can be measured both in vitro and in cells. AZ also inhibits cellular uptake of polyamines. This chapter comprises seven subchapters describing methods for studying expression and function of AZ.
authors.affiliation: Department of Molecular Biology, The Jikei University School of Medicine, Minato-ku, Tokyo, Japan.
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Murai
authors.forename: Yasuko
authors.initials: Y
authors.lastname: Murakami
authors.forename: Senya
authors.initials: S
authors.lastname: Matsufuji
mesh_terms: D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D015153:Blotting, Western; D002457:Cell Extracts; D045744:Cell Line, Tumor; D057075:Enzyme Assays / Q000379:methods*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D018965:Frameshifting, Ribosomal / Q000187:drug effects; D006801:Humans; D007150:Immunohistochemistry; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D007668:Kidney / Q000201:enzymology; D051379:Mice; D010766:Phosphorylation / Q000187:drug effects; D011073:Polyamines / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D011506:Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D014162:Transfection
chemical_list: D002457:Cell Extracts; D004791:Enzyme Inhibitors; D011073:Polyamines; D011506:Proteins; D011994:Recombinant Proteins; C031017:ornithine decarboxylase antizyme