title: In vitro folding, functional characterization, and disulfide pattern of the extracellular domain of human GLP-1 receptor.
abstract: The N-terminal, extracellular domain of the receptor for glucagon-like peptide 1 (GLP-1 receptor) was expressed at a high level in E. coli and isolated as inclusion bodies. Renaturation with concomitant disulfide bond formation was achieved from guanidinium-solubilized material. A soluble and active fraction of the protein was isolated by ion exchange chromatography and gel filtration. Complex formation with GLP-1 was shown by cross-linking experiments, surface plasmon resonance measurements, and isothermal titration calorimetry. The existence of disulfide bridges in the N-terminal receptor fragment was proven after digestion of the protein with pepsin. Further analysis revealed a disulfide-binding pattern with links between cysteines 46 and 71, 62 and 104, and between 85 and 126.
authors.affiliation: Institut für Biotechnologie der Martin-Luther-Universität Halle-Wittenberg, Halle, Germany.
authors.forename: Ariuna
authors.initials: A
authors.lastname: Bazarsuren
authors.forename: Ulla
authors.initials: U
authors.lastname: Grauschopf
authors.forename: Manfred
authors.initials: M
authors.lastname: Wozny
authors.forename: Dietmar
authors.initials: D
authors.lastname: Reusch
authors.forename: Eike
authors.initials: E
authors.lastname: Hoffmann
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Schaefer
authors.forename: Steffen
authors.initials: S
authors.lastname: Panzner
authors.forename: Rainer
authors.initials: R
authors.lastname: Rudolph
mesh_terms: D000595:Amino Acid Sequence; D002151:Calorimetry; D002942:Circular Dichroism; D004220:Disulfides / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005934:Glucagon / Q000378:metabolism; D052216:Glucagon-Like Peptide 1; D000067757:Glucagon-Like Peptide-1 Receptor; D006801:Humans; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism; D011485:Protein Binding; D017510:Protein Folding; D011498:Protein Precursors / Q000378:metabolism; D017434:Protein Structure, Tertiary; D018027:Receptors, Glucagon / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D017186:Titrimetry
chemical_list: D004220:Disulfides; C000593833:GLP1R protein, human; D000067757:Glucagon-Like Peptide-1 Receptor; D010446:Peptide Fragments; D011498:Protein Precursors; D018027:Receptors, Glucagon; D052216:Glucagon-Like Peptide 1; D005934:Glucagon