title: Baculovirus-driven expression and purification of glycine receptor alpha 1 homo-oligomers.
abstract: The glycine receptor is a ligand-gated anion channel protein of postsynaptic membranes. We expressed a homo-oligomeric receptor composed of human alpha 1 subunits in Spodoptera frugiperda cells by infection with a recombinant Autographa californica nuclear polyhedrosis virus. A substantial fraction of the recombinant receptor was incorporated as a functional channel protein into the cell's plasma membrane at expression levels 4- to 30-fold higher than in other eukaryotic heterologous expression systems or native rat spinal cord membranes, respectively. Upon detergent solubilization, the alpha 1 receptor was found to exist in a predominantly monodisperse state and could be affinity-purified to near homogeneity. This preparation is a potential starting point for future crystallisation studies.
authors.affiliation: Abteilung Neurochemie, Max-Planck-Institut für Hirnforschung, Frankfurt, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Morr
authors.forename: N
authors.initials: N
authors.lastname: Rundström
authors.forename: H
authors.initials: H
authors.lastname: Betz
authors.forename: D
authors.initials: D
authors.lastname: Langosch
authors.forename: B
authors.initials: B
authors.lastname: Schmitt
mesh_terms: D000818:Animals; D001483:Base Sequence; D001704:Biopolymers; D015153:Blotting, Western; D002460:Cell Line; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000235:genetics; D009838:Oligodeoxyribonucleotides; D018009:Receptors, Glycine / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D012995:Solubility; D018411:Spodoptera
chemical_list: D001704:Biopolymers; D009838:Oligodeoxyribonucleotides; D018009:Receptors, Glycine