Lundstrom Kenneth
PanTherapeutics, Lutry, Switzerland.
Methods Enzymol. 2015;556:331-50. doi: 10.1016/bs.mie.2014.11.047. Epub 2015 Mar 20.
Due to their importance as targets for drug development, rapid and consistent high-level production of G protein-coupled receptors (GPCRs) has become an essential part of drug discovery. Alphaviruses, particularly recombinant Semliki Forest virus (SFV) particles, have provided the means for expression of a number of GPCRs in a broad range of mammalian host cell lines for pharmacological characterization by determination of receptor binding activity and functional coupling to G proteins. The rapid high-titer virus particle production has made it possible to study a large number of GPCRs in parallel. Moreover, large-scale production in adherent and suspension cultures of mammalian cells has provided sufficient amounts of GPCRs for purification and subsequent structural studies. Furthermore, the high preference for neuronal delivery of SFV particles has allowed functional and localization studies of recombinant proteins in hippocampal slice cultures, in primary neurons, and in vivo.
由于作为药物开发靶点的重要性，快速且一致地高水平生产G蛋白偶联受体（GPCRs）已成为药物发现的重要组成部分。甲病毒，特别是重组Semliki森林病毒（SFV）颗粒，为在多种哺乳动物宿主细胞系中表达多种GPCRs提供了手段，以便通过测定受体结合活性和与G蛋白的功能偶联来进行药理学表征。快速的高滴度病毒颗粒生产使得能够并行研究大量的GPCRs。此外，在哺乳动物细胞的贴壁培养和悬浮培养中进行大规模生产，为纯化及后续的结构研究提供了足够量的GPCRs。此外，SFV颗粒对神经元递送的高度偏好，使得能够在海马切片培养物、原代神经元以及体内对重组蛋白进行功能和定位研究。
