Protocols for Studying Antizyme Expression and Function
Protocols for Studying Antizyme Expression and Function
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DOI：
10.1007/978-1-61779-034-8_15
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发表时间：
2011-01-01
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影响因子：
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通讯作者：
Matsufuji, Senya
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文献类型：
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作者：
Murai, Noriyuki;Murakami, Yasuko;Matsufuji, Senya
抗酶(AZ)是细胞内多胺反馈调节的关键分子。它是由多胺通过在其翻译过程中刺激核糖体移位而诱导的。在哺乳动物中，AZ被分成三个并列动物，AZ1-3。通过免疫化学方法或荧光标记蛋白的表达来确定，组织和亚细胞分布在不同的对虾中是不同的。只有AZ2已知是磷酸化的。AZ通过多种机制调节细胞内多胺水平。它与鸟氨酸脱羧酶(ODC)结合形成失活的复合体，并通过26S蛋白酶体对ODC进行降解。AZ促进ODC降解的活性可以在体外和细胞内测量。AZ还抑制细胞对多胺的摄取。这一章包括七个小节，描述了研究AZ的表达和功能的方法。
Antizyme (AZ) is a key molecule in feedback regulation of cellular polyamines. It is induced by polyamines through stimulation of ribosomal ftameshifting 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.