title: Prolonged signalling and trafficking of the bradykinin B2 receptor stimulated with the amphibian peptide maximakinin: insight into the endosomal inactivation of kinins.
abstract: Maximakinin, a 19-residue peptide from the amphibian Bombina maxima, incorporates the full sequence of bradykinin (BK) at its C-terminus with a hydrophilic 10-residue N-terminal extension. As a putative venom component, it may stimulate BK B(2) receptors (B(2)Rs) in a distinct manner relative to the fragile mammalian agonist BK. Maximakinin affinity for B(2)Rs and angiotensin converting enzyme (ACE) and its pharmacological profile have been compared to those of BK. Maximakinin is an agonist of the human and rabbit B(2)R with a 8-12 fold lesser potency, but a prolonged duration of action relative to BK (ERK MAP kinase activation, c-Fos induction in HEK 293 cells). Maximakinin had a moderately inferior affinity (∼6-fold vs. BK) for recombinant ACE based on [(3)H]enalaprilat binding displacement. Unlike BK, maximakinin induced the internalization of the fusion protein B(2)R-green fluorescent protein (GFP) and the downregulation of this construction over a 12-h stimulation period, reproducing the effect of inactivation-resistant B(2)R agonists. Alternate homologues of BK extended at the N-terminus showed intermediate behaviours between BK and maximakinin in the B(2)R-GFP downregulation assay. The recycling of B(2)R-GFP at the cell surface after a 3-h BK treatment was notably inhibited by cotreatment with E-64 or bafilomycin A1, supporting that an endosomal cysteine protease degrades kinins in a process that determines the cycling and fate of the B(2)R. Maximakinin is the first known natural kinin sequence that elicits a prolonged cellular signalling, thus suggesting a possible basis for a venomous action and a naturally selected one for the design of B(2)R-transported biotechnological cargoes.
authors: Centre de recherche en rhumatologie et immunologie, Centre Hospitalier Universitaire de Québec, Université Laval, CHUQ, 2705 Laurier Blvd., Québec, QC, Canada.
authors: Marie-Thérèse
authors: MT
authors: Bawolak
authors: Caroline
authors: C
authors: Roy
authors: Lajos
authors: L
authors: Gera
authors: François
authors: F
authors: Marceau
mesh_terms: D000664:Amphibian Venoms / Q000494:pharmacology*; D000663:Amphibians / Q000378:metabolism; D000818:Animals; D001920:Bradykinin / Q000378:metabolism*; D057056:Cysteine Proteases / Q000378:metabolism; D015536:Down-Regulation / Q000502:physiology; D011992:Endosomes / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D007930:Leucine / Q000031:analogs & derivatives / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000502:physiology; D018942:Macrolides / Q000378:metabolism; D020929:Mitogen-Activated Protein Kinase Kinases / Q000378:metabolism; D007703:Peptidyl-Dipeptidase A / Q000378:metabolism; D021381:Protein Transport; D016760:Proto-Oncogene Proteins c-fos / Q000378:metabolism; D011817:Rabbits; D043782:Receptor, Bradykinin B2 / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D000664:Amphibian Venoms; D018942:Macrolides; D016760:Proto-Oncogene Proteins c-fos; D043782:Receptor, Bradykinin B2; D011993:Recombinant Fusion Proteins; C497892:maximakinin, Bombina maxima; C040929:bafilomycin A1; D020929:Mitogen-Activated Protein Kinase Kinases; D057056:Cysteine Proteases; D007703:Peptidyl-Dipeptidase A; D007930:Leucine; C024974:E 64; D001920:Bradykinin