Overexpression of a glutamate receptor (GluR2) ligand binding domain in Escherichia coli: Application of a novel protein folding screen

摘要

Expression of the S1S2 ligand binding do- main [Kuusinen, A., Arvola, M. & Keinonen, K. (1995) EMBO J. 14, 6327-6332] of the rat α-amino-3-hydroxy-5-methylisox- azole-4-propionic acid-selective glutamate receptor GluR2 in Escherichia coli under control of a T7 promoter leads to production of >100 mg/liter of histidine-tagged S1S2 protein (HS1S2) in the form of inclusion bodies. Using a novel fractional factorial folding screen and a rational, step-by-step approach, multiple conditions were determined for the folding of the HS1S2 α-amino-3-hydroxy-5-methylisoxazole-4- propionic acid binding domain.