Bacteriophage membrane protein P9 as a fusion partner for the efficient expression of membrane proteins in Escherichia coli.
Despite their important roles and economic values, studies of membrane proteins have been hampered by the difficulties associated with obtaining sufficient amounts of protein. Here, we report a novel membrane protein expression system that uses the major envelope protein (P9) of phage φ6 as an N-terminal fusion partner. Phage membrane protein P9 facilitated the synthesis of target proteins and their integration into the Escherichia coli cell membrane. This system was used to produce various multi-pass transmembrane proteins, including G-protein-coupled receptors, transporters, and ion channels of human origin. Green fluorescent protein fusion was used to confirm the correct folding of the expressed proteins. Of the 14 membrane proteins tested, eight were highly expressed, three were moderately expressed, and three were barely expressed in E. coli. Seven of the eight highly expressed proteins could be purified after extraction with the mild detergent lauryldimethylamine-oxide. Although a few proteins have previously been developed as fusion partners to augment membrane protein production, we believe that the major envelope protein P9 described here is better suited to the efficient expression of eukaryotic transmembrane proteins in E. coli.
['Jung|Yuna|Y|', 'Jung|Hyeim|H|', 'Lim|Dongbin|D|']
D000595:Amino Acid Sequence D017126:Bacteriophage phi 6 D001483:Base Sequence D003001:Cloning, Molecular D003902:Detergents D004926:Escherichia coli D006801:Humans D007473:Ion Channels D008969:Molecular Sequence Data D010957:Plasmids D043562:Receptors, G-Protein-Coupled D011993:Recombinant Fusion Proteins D015854:Up-Regulation D014764:Viral Proteins
Membrane protein Fusion partner Phage φ6 G-protein-coupled receptor Membrane integration Therapeutic antibody