Nakane Shuhei, Matsuda Zene
China-Japan Joint Laboratory of Structural Virology and Immunology, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China.
Methods Mol Biol. 2015;1313:229-36. doi: 10.1007/978-1-4939-2703-6_17.
Fusion between viral and cellular membranes is the essential first step in infection of enveloped viruses. This step is mediated by viral envelope glycoproteins (Env) that recognize cellular receptors. The membrane fusion between the effector cells expressing viral Env and the target cells expressing its receptors can be monitored by several methods. We have recently developed a pair of chimeric reporter protein composed of split Renilla luciferase (RL) and split GFP. We named this reporter dual split protein (DSP), since it recovers both RL and GFP activities upon self reassociation. By using DSP, pore formation and content mixing between the effector and target cells can be monitored upon the recovery of RL and GFP activities after the membrane fusion. This quick assay provides quantitative as well as spatial information about membrane fusion mediated by viral Env.
病毒膜与细胞膜的融合是包膜病毒感染过程中至关重要的第一步。这一步骤由识别细胞受体的病毒包膜糖蛋白（Env）介导。表达病毒Env的效应细胞与表达其受体的靶细胞之间的膜融合可以通过多种方法进行监测。我们最近开发了一对由分裂型海肾荧光素酶（RL）和分裂型绿色荧光蛋白组成的嵌合报告蛋白。我们将这种报告蛋白命名为双分裂蛋白（DSP），因为它在自我重新结合后可恢复RL和GFP的活性。通过使用DSP，在膜融合后RL和GFP活性恢复时，可以监测效应细胞和靶细胞之间的孔形成和内容物混合情况。这种快速检测方法提供了有关病毒Env介导的膜融合的定量和空间信息。
