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Polyamines pp 237-267 | Cite as

Protocols for Studying Antizyme Expression and Function

  • Noriyuki Murai
  • Yasuko Murakami
  • Senya Matsufuji
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 720)

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.

Key words

Antizyme Ornithine decarboxylase Antizyme inhibitor Translational frameshifting Antibody Immunochemistry Localization Phosphorylation Protein degradation Polyamine transport 

Notes

Acknowledgment

The authors thank Dr. Jun-ichi Suzuki for providing detailed protocols.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Noriyuki Murai
    • 1
  • Yasuko Murakami
    • 1
  • Senya Matsufuji
    • 1
  1. 1.Department of Molecular BiologyThe Jikei University School of MedicineMinato-kuJapan

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