title: Structural basis of proline-specific exopeptidase activity as observed in human dipeptidyl peptidase-IV.
abstract: Inhibition of dipeptidyl peptidase IV (DPP-IV), the main glucagon-like peptide 1 (GLP1)-degrading enzyme, has been proposed for the treatment of type II diabetes. We expressed and purified the ectodomain of human DPP-IV in Pichia pastoris and determined the X-ray structure at 2.1 A resolution. The enzyme consists of two domains, the catalytic domain, with an alpha/beta hydrolase fold, and a beta propeller domain with an 8-fold repeat of a four-strand beta sheet motif. The beta propeller domain contributes two important functions to the molecule that have not been reported for such structures, an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif. The Glu motif provides recognition and a binding site for the N terminus of the substrates, as revealed by the complex structure with diprotin A, a substrate with low turnover that is trapped in the tetrahedral intermediate of the reaction in the crystal.
authors.affiliation: F. Hoffmann-La Roche AG, Pharma Research Discovery, 4070 Basel, Switzerland.
authors.forename: Ralf
authors.initials: R
authors.lastname: Thoma
authors.forename: Bernd
authors.initials: B
authors.lastname: Löffler
authors.forename: Martine
authors.initials: M
authors.lastname: Stihle
authors.forename: Walter
authors.initials: W
authors.lastname: Huber
authors.forename: Armin
authors.initials: A
authors.lastname: Ruf
authors.forename: Michael
authors.initials: M
authors.lastname: Hennig
mesh_terms: D000243:Adenosine Deaminase / Q000378:metabolism; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001665:Binding Sites; D018360:Crystallography, X-Ray; D019281:Dimerization; D018819:Dipeptidyl Peptidase 4 / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D004795:Enzyme Stability; D020689:Exopeptidases / Q000378:metabolism*; D006031:Glycosylation; D006801:Humans; D008024:Ligands; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000037:antagonists & inhibitors; D010843:Pichia / Q000201:enzymology; D011392:Proline; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013379:Substrate Specificity; D014867:Water / Q000737:chemistry
chemical_list: D008024:Ligands; D009842:Oligopeptides; D014867:Water; C041445:diprotin A; D011392:Proline; D020689:Exopeptidases; D018819:Dipeptidyl Peptidase 4; D000243:Adenosine Deaminase