title: Effects of vasopressin-mastoparan chimeric peptides on insulin release and G-protein activity.
abstract: Two chimeric peptides, consisting of the linear vasopressin receptor V1 antagonist PhAc-D-Tyr(Me)-Phe-Gln-Asn-Arg-Pro-Arg-Tyr, in the N-terminus and mastoparan in the C-terminus connected directly (M375) or via 6-aminohexanoic acid (M391), have been synthesised. At 10 microM concentration, these novel peptides increased insulin secretion from isolated rat pancreatic islet cells 18-26-fold at 3.3 mM glucose and 3.5-5-fold at 16.7 mM glucose. PTX pretreatment of the islets decreased the peptide-induced insulin release. M375 and M391 bind to V1a vasopressin receptors with affinities lower than the unmodified vasopressin antagonist, but with K(D) values of 3.76 nM and 9.02 nM, respectively, both chimeras are high affinity ligands. The GTPase activity and GTPgammaS binding in the presence of these peptides has been characterised in Rin m5F cells. Comparison of the influence of the peptides M375 and M391 on GTPase activity in native and pertussis toxin-treated cells suggests a selective activation of G alpha(i)/G alpha(o) subunits, combined with a suppression of other GTPases, primarily G alpha(s). However, the GTPgammaS binding data show that the peptides retain some of the activating property even in PTX-treated cell membranes. In conclusion, the conjugation of mastoparan with the V1a receptor antagonists produce peptides with properties different from the parent peptides that could be used to elucidate the role of different G proteins in insulin release.
authors.affiliation: Department of Neurochemistry and Neurotoxicology, Arrheniuslaboratories, Stockholm University, Sweden.
authors.forename: M
authors.initials: M
authors.lastname: Hällbrink
authors.forename: K
authors.initials: K
authors.lastname: Saar
authors.forename: C G
authors.initials: CG
authors.lastname: Ostenson
authors.forename: U
authors.initials: U
authors.lastname: Soomets
authors.forename: S
authors.initials: S
authors.lastname: Efendic
authors.forename: J
authors.initials: J
authors.lastname: Howl
authors.forename: M
authors.initials: M
authors.lastname: Wheatley
authors.forename: M
authors.initials: M
authors.lastname: Zorko
authors.forename: U
authors.initials: U
authors.lastname: Langel
mesh_terms: D000818:Animals; D065092:Antidiuretic Hormone Receptor Antagonists; D002462:Cell Membrane / Q000378:metabolism; D020558:GTP Phosphohydrolases / Q000187:drug effects / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000187:drug effects / Q000378:metabolism*; D005947:Glucose / Q000494:pharmacology; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D066298:In Vitro Techniques; D007328:Insulin / Q000378:metabolism*; D036341:Intercellular Signaling Peptides and Proteins; D007515:Islets of Langerhans / Q000187:drug effects / Q000378:metabolism; D008297:Male; D010455:Peptides / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology; D037342:Pertussis Toxin; D011485:Protein Binding; D051381:Rats; D017208:Rats, Wistar; D017483:Receptors, Vasopressin / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000494:pharmacology*; D014407:Tumor Cells, Cultured / Q000187:drug effects; D014667:Vasopressins / Q000378:metabolism / Q000494:pharmacology; D010566:Virulence Factors, Bordetella / Q000494:pharmacology; D014862:Wasp Venoms / Q000378:metabolism / Q000494:pharmacology*
chemical_list: D065092:Antidiuretic Hormone Receptor Antagonists; D007328:Insulin; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; D017483:Receptors, Vasopressin; D011993:Recombinant Fusion Proteins; D010566:Virulence Factors, Bordetella; D014862:Wasp Venoms; D014667:Vasopressins; D016244:Guanosine 5'-O-(3-Thiotriphosphate); C037500:mastoparan; D037342:Pertussis Toxin; D020558:GTP Phosphohydrolases; D019204:GTP-Binding Proteins; D005947:Glucose