title: Mambalgin-1 Pain-relieving Peptide, Stepwise Solid-phase Synthesis, Crystal Structure, and Functional Domain for Acid-sensing Ion Channel 1a Inhibition.
abstract: Mambalgins are peptides isolated from mamba venom that specifically inhibit a set of acid-sensing ion channels (ASICs) to relieve pain. We show here the first full stepwise solid phase peptide synthesis of mambalgin-1 and confirm the biological activity of the synthetic toxin both in vitro and in vivo. We also report the determination of its three-dimensional crystal structure showing differences with previously described NMR structures. Finally, the functional domain by which the toxin inhibits ASIC1a channels was identified in its loop II and more precisely in the face containing Phe-27, Leu-32, and Leu-34 residues. Moreover, proximity between Leu-32 in mambalgin-1 and Phe-350 in rASIC1a was proposed from double mutant cycle analysis. These data provide information on the structure and on the pharmacophore for ASIC channel inhibition by mambalgins that could have therapeutic value against pain and probably other neurological disorders.
authors.affiliation: From the Commissariat à l'Energie Atomique, iBiTecS, Service d'Ingénierie Moléculaire des Protéines, 91191 Gif-sur-Yvette.
authors.forename: Gilles
authors.initials: G
authors.lastname: Mourier
authors.affiliation: the CNRS, Institut de Pharmacologie Moléculaire et Cellulaire, the Université de Nice Sophia Antipolis, and the LabEx Ion Channel Science and Therapeutics, UMR 7275, 06560 Valbonne, France.
authors.forename: Miguel
authors.initials: M
authors.lastname: Salinas
authors.affiliation: From the Commissariat à l'Energie Atomique, iBiTecS, Service d'Ingénierie Moléculaire des Protéines, 91191 Gif-sur-Yvette.
authors.forename: Pascal
authors.initials: P
authors.lastname: Kessler
authors.affiliation: From the Commissariat à l'Energie Atomique, iBiTecS, Service d'Ingénierie Moléculaire des Protéines, 91191 Gif-sur-Yvette.
authors.forename: Enrico A
authors.initials: EA
authors.lastname: Stura
authors.affiliation: From the Commissariat à l'Energie Atomique, iBiTecS, Service d'Ingénierie Moléculaire des Protéines, 91191 Gif-sur-Yvette.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Leblanc
authors.affiliation: From the Commissariat à l'Energie Atomique, iBiTecS, Service d'Ingénierie Moléculaire des Protéines, 91191 Gif-sur-Yvette.
authors.forename: Livia
authors.initials: L
authors.lastname: Tepshi
authors.affiliation: the CNRS, Institut de Pharmacologie Moléculaire et Cellulaire, the Université de Nice Sophia Antipolis, and the LabEx Ion Channel Science and Therapeutics, UMR 7275, 06560 Valbonne, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Besson
authors.affiliation: the CNRS, Institut de Pharmacologie Moléculaire et Cellulaire, the Université de Nice Sophia Antipolis, and the LabEx Ion Channel Science and Therapeutics, UMR 7275, 06560 Valbonne, France.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Diochot
authors.affiliation: the CNRS, Institut de Pharmacologie Moléculaire et Cellulaire, the Université de Nice Sophia Antipolis, and the LabEx Ion Channel Science and Therapeutics, UMR 7275, 06560 Valbonne, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Baron
authors.affiliation: the CNRS, Institut de Pharmacologie Moléculaire et Cellulaire, the Université de Nice Sophia Antipolis, and.
authors.forename: Dominique
authors.initials: D
authors.lastname: Douguet
authors.affiliation: the CNRS, Institut de Pharmacologie Moléculaire et Cellulaire, the Université de Nice Sophia Antipolis, and the LabEx Ion Channel Science and Therapeutics, UMR 7275, 06560 Valbonne, France lingueglia@ipmc.cnrs.fr.
authors.forename: Eric
authors.initials: E
authors.lastname: Lingueglia
authors.affiliation: From the Commissariat à l'Energie Atomique, iBiTecS, Service d'Ingénierie Moléculaire des Protéines, 91191 Gif-sur-Yvette, denis.servent@cea.fr.
authors.forename: Denis
authors.initials: D
authors.lastname: Servent
mesh_terms: D062565:Acid Sensing Ion Channels / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D004546:Elapid Venoms / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology; D019906:Nuclear Magnetic Resonance, Biomolecular; D009865:Oocytes; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D051381:Rats; D014982:Xenopus laevis
chemical_list: D062565:Acid Sensing Ion Channels; C568268:Asic1 protein, rat; D004546:Elapid Venoms; D010455:Peptides; C576673:mambalgin-1, Dendroaspis polylepis
keywords: X-ray crystallography; acid sensing ion channel (ASIC); mambalgin; pain; peptide chemical synthesis; peptide interaction; sodium channel; structural model; toxin