Expression of Fully Functional Gpcr in E. Coli

One-third of eukaryotic proteins are associated with membranes and these membrane proteins (MP) represent approximately 50% of pharmacological targets. Researchers are therefore nursing the hope of achieving a higher percentage of about 80% in the near future. The only bottleneck to achieving this target is the experimental challengers associated with the production, purification and crystallization at reasonable cost. Expression of protein in E. coli is now very popular, because it is easy to use and also comes with low operational cost. But membrane proteins have been difficult to produce in E. coli. The membrane proteins (MP), G-protein coupled receptors (GPCRs) represent the major class of potential drug targets but probably the most difficult class of MPs to bring to three-dimensional studies. This review paper therefore sought to bring to the fore some of the techniques employed by researchers to improve the quality and quantity of GPCR expressed in E. coli. A literature search on the effective expression of some important members of the GPCR family in E. coli was done. The Expression of neurotensin receptor type1 (NTIS1) in BL21 (DE3) and C41 (DE3) strains of E. coli; Autoinduction of NTS1 in BL21 (DE3) with the medium MagicmediaTM; Expression in E. coli by targeting the inner membrane with fusion partners MBP and TRX (MBP-GPCR-TRX); Expression in E. coli by targeting the inclusion bodies with fusion partners GST and TRX; and the use of pDEST17 vectors and C43 strain at 37 o C have proved experimentally efficient to express highly functional GPCR. INTRODUCTION: It is estimated that one-third of eukaryotic protein are associated with membranes 1 , and these proteins (membrane proteins) have been established to represent approximately 50% of pharmacological targets 2 . QUICK RESPONSE CODE IJPSR: ICV (2011)5.07 Article can be accessed online on: www.ijpsr.com It is the hope of all researchers in the field of drug discovery that, this number will increase up to 80% of all new drug targets for the next decade 2 . The only difficulty to achieving the above target is the fact that, there are experimental challenges associated with the production, purification and crystallization of membrane proteins (MP). Membrane proteins, especially those from eukaryotes, have been difficult to produce in E. coli and therefore have limited the ability to use this inexpensive and robust host to produce protein for biochemical and structural studies 3 .