title: Identification of a pepducin acting as S1P3 receptor antagonist.
abstract: Sphingosine-1-phosphate (S1P) is a bioactive lipid with key functions in the immune, inflammatory, and cardiovascular systems. S1P exerts its action through the interaction with a family of five known G protein-coupled receptors, named S1P(1-5). Among them, S1P(3) has been implicated in the pathological processes of a number of diseases, including sepsis and cancer. KRX-725 (compound 1) is a pepducin that mimics the effects of S1P by triggering specifically S1P(3). Here, aiming to identify novel S1P(3) antagonists, we carried out an alanine scanning analysis to address the contribution of the side chains of each amino acid residue to the peptide function. Then, deleted peptides from both the C- and N-terminus were prepared in order to determine the minimal sequence for activity and to identify the structural requirements for agonistic and, possibly, antagonistic behaviors. The pharmacological results of the Ala-scan derived compounds (2-10) suggested a high tolerance of the pepducin 1 to amino acid substitutions. Importantly, the deleted peptide 16 has the ability to inhibit, in a dose-dependent manner, both pepducin 1-induced vasorelaxation and fibroblast proliferation. Finally, a computational analysis was performed on the prepared compounds, showing that the supposed antagonists 16 and 17 appeared to be aligned with each other but not with the others. These results suggested a correlation between specific conformations and activities.
authors.affiliation: Dipartimento di Farmacia, Università di Napoli 'Federico II', Via D. Montesano, 49, 80131, Napoli, Italy.
authors.forename: Beatrice
authors.initials: B
authors.lastname: Severino
authors.forename: Giuseppina Maria
authors.initials: GM
authors.lastname: Incisivo
authors.forename: Ferdinando
authors.initials: F
authors.lastname: Fiorino
authors.forename: Antonio
authors.initials: A
authors.lastname: Bertolino
authors.forename: Francesco
authors.initials: F
authors.lastname: Frecentese
authors.forename: Francesco
authors.initials: F
authors.lastname: Barbato
authors.forename: Serena
authors.initials: S
authors.lastname: Manganelli
authors.forename: Giada
authors.initials: G
authors.lastname: Maggioni
authors.forename: Domenica
authors.initials: D
authors.lastname: Capasso
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Caliendo
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Santagada
authors.forename: Raffaella
authors.initials: R
authors.lastname: Sorrentino
authors.forename: Fiorentina
authors.initials: F
authors.lastname: Roviezzo
authors.forename: Elisa
authors.initials: E
authors.lastname: Perissutti
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001013:Aorta, Thoracic / Q000187:drug effects / Q000502:physiology; D049109:Cell Proliferation / Q000187:drug effects; D057846:Cell-Penetrating Peptides / Q000737:chemistry / Q000494:pharmacology*; D002478:Cells, Cultured; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D008958:Models, Molecular; D009119:Muscle Contraction / Q000187:drug effects; D010446:Peptide Fragments / Q000737:chemistry / Q000494:pharmacology*; D049349:Receptors, Lysosphingolipid / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D000081025:Sphingosine-1-Phosphate Receptors; D014665:Vasodilator Agents / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D057846:Cell-Penetrating Peptides; D010446:Peptide Fragments; D049349:Receptors, Lysosphingolipid; C501285:S1pr3 protein, mouse; D000081025:Sphingosine-1-Phosphate Receptors; D014665:Vasodilator Agents; C000588569:myristoyl-glycyl-methionyl-arginyl-prolyl-tyrosyl-aspartyl-alanyl-asparagyl-lysyl-argininamide
keywords: Ala-scan; S1P3 antagonist; pepducin; sphingosine-1-phosphate (S1P)