title: Baculovirus-mediated expression of GPCRs in insect cells.
abstract: G-protein-coupled receptors (GPCRs) are a large family of seven transmembrane proteins that influence a considerable number of cellular events. For this reason, they are one of the most studied receptor types for their pharmacological and structural properties. Solving the structure of several GPCR receptor types has been possible using almost all expression systems, including Escherichia coli, yeast, mammalian, and insect cells. So far, however, most of the GPCR structures solved have been done using the baculovirus insect cell expression system. The reason for this is mainly due to cost-effectiveness, posttranslational modification efficiency, and overall effortless maintenance. The system has evolved so much that variables starting from vector type, purification tags, cell line, and growth conditions can be varied and optimized countless ways to suit the needs of new constructs. Here, we present the array of techniques that enable the rapid and efficient optimization of expression steps for maximal protein quality and quantity, including our emendations.
authors: Department of Biochemistry, Helsinki University, Helsinki, Finland.
authors: Tuulia
authors: T
authors: Saarenpää
authors: Novartis Institutes of Biomedical Research, Basel, Switzerland.
authors: Veli-Pekka
authors: VP
authors: Jaakola
authors: Department of Biochemistry, Helsinki University, Helsinki, Finland; School of Biomedical Sciences, Faculty of Biological Sciences, University of Leeds, Leeds, United Kingdom. Electronic address: adrian.goldman@helsinki.fi.
authors: Adrian
authors: A
authors: Goldman
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D019149:Bioreactors; D002460:Cell Line; D004274:DNA, Recombinant / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007313:Insecta / Q000166:cytology / Q000235:genetics*; D008958:Models, Molecular; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry / Q000235:genetics*; D014162:Transfection / Q000379:methods*
chemical_list: D004274:DNA, Recombinant; D043562:Receptors, G-Protein-Coupled
keywords: Baculovirus; GPCR; High-throughput; Insect cell; Large-scale insect cell expression; WAVE bioreactor