Efetova Marina, Schwärzel Martin
Institute for Biology/Neurobiology, Freie Universität Berlin, Königin-Luise-Str. 28-30, Berlin, D-14195, Germany.
Methods Mol Biol. 2015;1294:131-5. doi: 10.1007/978-1-4939-2537-7_10.
Photoactivatable adenylyl cyclases (PACs) are proteins that combine the capacity of a photoreceptor with that of an adenylyl cyclase. When ectopically expressed under the control of specific promoters, these naturally occurring proteins become potent transgenic tools that facilitate the increase of cellular cAMP levels by the use of light. Currently, three different PAC transgenes-the euglenoid euPACα and euPACβ, as well as the b eggiatoan bPac-are available. These transgenic tools provide cyclase activity capable of increasing cellular cAMP levels up to a hundredfold with either phasic- or tonic-like kinetic characteristics. Here, we consider the functional features of different cyclases and provide operating guidelines to optimize the use of PACs in vivo.
光激活腺苷酸环化酶(PACs)是一种将光感受器的功能与腺苷酸环化酶的功能结合在一起的蛋白质。当在特定启动子的控制下异位表达时,这些天然存在的蛋白质成为强大的转基因工具,可通过光照来提高细胞内cAMP水平。目前,有三种不同的PAC转基因——眼虫类的euPACα和euPACβ,以及贝氏硫细菌的bPac。这些转基因工具提供的环化酶活性能够将细胞内cAMP水平提高多达百倍,具有类似相位或紧张性的动力学特征。在这里,我们考虑了不同环化酶的功能特性,并提供了操作指南,以优化PACs在体内的使用。