BACULOVIRUS-DRIVEN EXPRESSION AND PURIFICATION OF GLYCINE RECEPTOR ALPHA-L HOMO-OLIGOMERS

The glycine receptor is a ligand-gated anion channel protein of postsy naptic membranes. We expressed a homo-oligomeric receptor composed of human al subunits in Spodoptera frugiperda cells by infection with a r ecombinant Autographa californica nuclear polyhedrosis virus. A substa ntial fraction of the recombinant receptor was incorporated as a funct ional channel protein into the cell's plasma membrane at expression le vels 4- to 30-fold higher than in other eukaryotic heterologous expres sion systems or native rat spinal cord membranes, respectively, Upon d etergent solubilization, the alpha 1 receptor was found to exist in a predominantly monodisperse state and could be affinity-purified to nea r homogeneity, This preparation is a potential starting point for futu re crystallisation studies.