公开(公告)号：EP3194960A1

公开(公告)日：2017-07-26

申请号：EP15841561.2

申请日：2015-09-21

申请人： The Royal Institution for the Advancement of Learning / McGill University ,  Université de Montréal

发明人： LAPORTE, Stéphane Alain ,  NAMKUNG, Yoon ,  BOUVIER, Michel ,  LE GOUILL, Christian ,  HOGUE, Mireille ,  LUKASHEVA, Viktoriya ,  KOBAYASHI, Hiroyuki ,  DEBLOIS, Denis ,  DURETTE, Etienne

IPC分类号： G01N33/53 ,  C12M1/34 ,  C12N15/62 ,  C12Q1/02 ,  C12Q1/66 ,  G01N33/58

CPC分类号： C07K14/723 ,  C07K2319/60 ,  G01N33/5035

摘要： Bioluminescence resonance energy transfer (BRET) biosensors for assessing the intracellular localization, internalization and trafficking into cellular compartments of proteins such as receptors, and other biomolecules such as second messengers, are disclosed. These biosensors, which are dependent on the concentration/density of the BRET donor and acceptor in cellular compartments rather that specific protein-protein interactions, use a Renilla GFP/Luc BRET pair, which allows the robust and reproducible monitoring of protein trafficking/localization, with a sensitivity compatible with high-throughput screening (HTS). The use of these biosensors for various applications, including assessing/monitoring protein endocytosis, recycling and intra-cellular trafficking, receptor maturation/rescue by pharmacological chaperones, various endocytosis/exocytosis processes, activation/inhibition, as well as biomolecule concentration/density in different cellular compartments, is also disclosed.

摘要翻译： 公开了生物发光共振能量转移（BRET）生物传感器，用于评估细胞内定位，内化和运输到蛋白质如受体和其他生物分子如第二信使的细胞区室中。 这些生物传感器依赖于细胞区室中BRET供体和受体的浓度/密度而非特定的蛋白质 - 蛋白质相互作用，使用Renilla GFP / Luc BRET对，其允许对蛋白质运输/定位的稳健和可重现的监测， 灵敏度与高通量筛选（HTS）兼容。 将这些生物传感器用于各种应用，包括评估/监测蛋白质内吞作用，再循环和细胞内运输，通过药理学分子伴侣的受体成熟/拯救，各种内吞作用/胞吐作用过程，活化/抑制以及不同细胞区室中的生物分子浓度/密度 ，也被披露。