| Title: | An efficient Escherichia coli expression system for the production of a functional N-terminal domain of the T1R3 taste receptor |
| Author(s): | Maitrepierre E; Sigoillot M; Le Pessot L; Briand L; |
| Address: | "Centre des Sciences du Gout et de l'Alimentation, UMR-1324 INRA, UMR-6265 CNRS, Universite de Bourgogne, 21000 Dijon, France" |
| ISSN/ISBN: | 2165-5987 (Electronic) 2165-5979 (Print) 2165-5979 (Linking) |
| Abstract: | "Sweet taste is mediated by a dimeric receptor composed of two distinct subunits, T1R2 and T1R3, whereas the T1R1/T1R3 receptor is involved in umami taste perception. The T1R1, T1R2, and T1R3 subunits are members of the small family of class C G protein-coupled receptors (GPCRs). The members of this family are characterized by a large N-terminal domain (NTD), which is structurally similar to bacterial periplasmic-binding proteins and contains the primary ligand-binding site. In a recent study, we described a strategy to produce a functional dimeric human T1R3-NTD. Although the protein was expressed as inclusion bodies (IBs) using the Escherichia coli system, the conditions for the refolding of functional hT1R3-NTD were determined using a fractional factorial screen coupled to a binding assay. Here, we report that this refolding strategy can be used to produce T1R1- and T1R2-NTDs in large quantities. We also discuss that our findings could be more generally applicable to other class C GPCR-NTDs, including the gamma-aminobutyric acid type B receptor (GABABR), the extracellular calcium-sensing receptor (CaSR) and the large family of pheromone (V2R) orphan receptors" |
| Keywords: | Escherichia coli Gpcr bacteria expression recombinant protein sugar sweet receptor sweetener taste umami receptor; |
| Notes: | "PubMed-not-MEDLINEMaitrepierre, Elodie Sigoillot, Maud Le Pessot, Laurence Briand, Loic eng Comment Research Support, Non-U.S. Gov't 2012/08/23 Bioengineered. 2013 Jan-Feb; 4(1):25-9. doi: 10.4161/bioe.21877. Epub 2012 Aug 22" |
