Roberts Thomas H, Ahn Joon-Woo, Lampl Nardy, Fluhr Robert
Department of Chemistry and Biomolecular Sciences, Macquarie University, North Ryde, Australia.
Methods Enzymol. 2011;499:347-66. doi: 10.1016/B978-0-12-386471-0.00017-1.
Serpins appear to be ubiquitous in the Plant Kingdom and have several unique properties when compared to the substantial number of other families of protease inhibitors in plants. Serpins in plants are likely to have functions distinct from those of animal serpins, partly because plants and animals developed multicellularity independently and partly because most animal serpins are involved in animal-specific processes, such as blood coagulation and the activation of complement. To encourage and facilitate the discovery of plant serpin functions, here we provide a set of protocols for detection of serpins in plant extracts, localization of serpins in plant tissues and cells, purification of serpins from a range of organs from monocot and eudicot plants, production and purification of recombinant plant serpins, and analysis of plant-protease interactions including identification of in vivo target proteases.
丝氨酸蛋白酶抑制剂（Serpins）似乎在植物界无处不在，与植物中大量其他蛋白酶抑制剂家族相比，具有几个独特的特性。植物中的丝氨酸蛋白酶抑制剂可能具有与动物丝氨酸蛋白酶抑制剂不同的功能，部分原因是植物和动物独立地发展出多细胞性，部分原因是大多数动物丝氨酸蛋白酶抑制剂参与动物特有的过程，如血液凝固和补体激活。为了鼓励和促进植物丝氨酸蛋白酶抑制剂功能的发现，我们在此提供一套方案，用于检测植物提取物中的丝氨酸蛋白酶抑制剂、在植物组织和细胞中定位丝氨酸蛋白酶抑制剂、从单子叶植物和双子叶植物的一系列器官中纯化丝氨酸蛋白酶抑制剂、生产和纯化重组植物丝氨酸蛋白酶抑制剂，以及分析植物与蛋白酶的相互作用，包括鉴定体内靶蛋白酶。
