| 摘要 |
<p>The invention relates to chemical synthesis of extramembranous receptor domains of membrane protein receptors, and compositions and methods that employ them. The extramembranous receptor domains include the soluble ligand-binding extracellular and cytosolic domains of membrane protein receptors. The extramembranous receptor domains of the invention are produced by ligating under chemoselective chemical ligation conditions first and second peptides of an extramembranous receptor domain of a membrane protein receptor, where the peptides have unprotected chemoselective reactive groups capable of forming a covalent bond therein between. The ligation product is exposed to a folding buffer having a chaotropic reagent and an organic solvent that approximates the water-lipid interface of a cell membrane. Exposure to the folding buffer is followed by isolation from the buffer of ligation product that binds to a ligand of the membrane protein receptor. The ligand-binding portion of the ligation product produced by this method represents folded extramembranous receptor domain. The invention is exemplified by total chemical synthesis of the N-terminal domain of the glucagon-like peptide 1 receptor, demonstration of its ability to bind to peptide ligand and characterization of its disulfide map.</p> |
