研究抗酶表达与功能的实验方案。

Protocols for studying antizyme expression and function.

作者信息

Murai Noriyuki, Murakami Yasuko, Matsufuji Senya

机构信息

Department of Molecular Biology, The Jikei University School of Medicine, Minato-ku, Tokyo, Japan.

出版信息

Methods Mol Biol. 2011;720:237-67. doi: 10.1007/978-1-61779-034-8_15.

DOI:10.1007/978-1-61779-034-8_15

PMID:21318878

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.

摘要

抗酶（AZ）是细胞多胺反馈调节中的关键分子。它在翻译过程中通过核糖体移码的刺激由多胺诱导产生。在哺乳动物中，AZ分化为三个旁系同源物，即AZ1 - 3。通过免疫化学方法或荧光标记蛋白的表达确定，这些旁系同源物在组织和亚细胞分布上有所不同。已知只有AZ2会被磷酸化。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）结合形成无活性复合物，并触发26S蛋白酶体对ODC的降解。促进ODC降解的AZ活性可以在体外和细胞中进行测量。AZ还抑制细胞对多胺的摄取。本章包括七个小节，描述了研究AZ表达和功能的方法。