title: A peptide mimic of an antigenic loop of alpha-human chorionic gonadotropin hormone: solution structure and interaction with a llama V(HH) domain.
abstract: The X-ray structure of a ternary complex between human chorionic gonadotropin hormone (hCG) and two Fvs recognizing its alpha and beta subunits has been recently determined. The Fvs recognize the elongated hCG molecule by its two ends, one being the Leu-12-Cys-29 loop of the alpha subunit. We have designed and synthesized a 17-amino-acid peptide (named PepH14) derived from the sequence of this antigenic loop with the purpose of mimicking its three-dimensional structure and its affinity for antibodies. We have determined the solution structure of PepH14 by homonuclear NMR spectroscopy and derived distance restraints. Comparison of this structure with that of the corresponding antigenic loop of alpha-hCG reveals strong conformational similarities. In particular, the two pairs of residues that establish crucial contacts with the Fv fragment share the same conformation in PepH14 and in the authentic hormone loop. We propose a three-dimensional model of interaction of PepH14 with a llama V(HH) (V(HH)-H14) fragment cloned from a single-chain llama immunoglobulin raised against alpha-hCG. This model has been constrained by the chemical shift variations of the H14 1HN and 15N resonances monitored upon binding with PepH14. Mapping of the backbone chemical shift variations on the V(HH) structure determined by NMR indicates that PepH14 binds to V(HH)-H14 and forms a complex using the three complementary determining regions (CDRs). They define a shallow groove encompassing residues Thr-31, Ala-56, Tyr-59 and Trp-104 which have been shown to be in conformational exchange [Renisio, Pérez, Czisch, Guenneugues, Bornet, Frenken, Cambillau and Darbon (2002) Proteins 47, 546-555] and also Phe-37 and Ala-50. This groove is close to the hydrophobic interface area observed between VH and VL domains in Fvs from classical antibodies, which explains the rather lateral binding of PepH14 on the V(HH).
authors: Architecture et Fonction des Macromolécules Biologiques, UMR 6098, CNRS and Universités d'Aix-Marseille I and II, 31 Chemin Joseph Aiguier, 13402 Marseille, CEDEX 20, France.
authors: Gilles
authors: G
authors: Ferrat
authors: Jean-Guillaume
authors: JG
authors: Renisio
authors: Xavier
authors: X
authors: Morelli
authors: Jerry
authors: J
authors: Slootstra
authors: Rob
authors: R
authors: Meloen
authors: Christian
authors: C
authors: Cambillau
authors: Hervé
authors: H
authors: Darbon
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002161:Camelids, New World; D015292:Glycoprotein Hormones, alpha Subunit / Q000737:chemistry*; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry*; D011487:Protein Conformation; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
chemical_list: D015292:Glycoprotein Hormones, alpha Subunit; D007135:Immunoglobulin Variable Region; D010446:Peptide Fragments