title: A novel bradykinin potentiating peptide isolated from Bothrops jararacussu venom using catallytically inactive oligopeptidase EP24.15.
abstract: Characterization of the peptide content of venoms has a number of potential benefits for basic research, clinical diagnosis, development of new therapeutic agents, and production of antiserum. Here, we use a substrate-capture assay that employs a catalytically inactive mutant of thimet oligopeptidase (EC 3.4.24.15; EP24.15) to identify novel bioactive peptides in Bothrops jararacussu venom. Of the peptides captured with inactive EP24.15 and identified by mass spectrometry, three were previously identified bradykinin-potentiating peptides (BPP), <ENWPHPQIPP (Xc), <EGGWPRPGPEIPP (XIIIa) and <EARPPHPPIPP (XIe) (where <E is a pyroglutamyl residue). In addition, we identified a novel BPP peptide containing additional AP amino acids in the C-terminus (<EARPPHPPIPPAP); this novel peptide was named BPP-AP. Next, dermal and muscle microcirculations were visualized using intravital microscopy to establish the roles of peptides BPP-XIe and BPP-AP in this process. After local administration of peptide BPP-XIe (0.5 microg.microL(-1)), leukocyte rolling flux and adhesion were increased by fivefold in post-capillary venules, without any increments in vasodilatation of arterioles compared to control experiments. In contrast, local administration of BPP-AP (0.5 microg.microL(-1)) potently induced vasodilatation of arterioles (nearly 100% increase compared with the vehicle saline control), with only a small increase in leukocyte rolling flux. Therefore, the novel BPP-AP described herein has pharmacological advantages compared to the BPP-XIe. The present study further suggests that inactive oligopeptidase EP24.15 is a useful tool for the isolation of bioactive peptides from crude biological samples.
authors.affiliation: Laboratório Especial de Toxinologia Aplicada-CAT/CEPID, Instituto Butantan, São Paulo, Brazil. vrioli@butantan.gov.br
authors.forename: Vanessa
authors.initials: V
authors.lastname: Rioli
authors.forename: Benedito C
authors.initials: BC
authors.lastname: Prezoto
authors.forename: Katsuhiro
authors.initials: K
authors.lastname: Konno
authors.forename: Robson L
authors.initials: RL
authors.lastname: Melo
authors.forename: Clécio F
authors.initials: CF
authors.lastname: Klitzke
authors.forename: Emer S
authors.initials: ES
authors.lastname: Ferro
authors.forename: Mônica
authors.initials: M
authors.lastname: Ferreira-Lopes
authors.forename: Antonio C M
authors.initials: AC
authors.lastname: Camargo
authors.forename: Fernanda C V
authors.initials: FC
authors.lastname: Portaro
mesh_terms: D000595:Amino Acid Sequence; D000806:Angiotensin-Converting Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001794:Blood Pressure / Q000187:drug effects; D017837:Bothrops; D001920:Bradykinin / Q000378:metabolism* / Q000494:pharmacology; D003435:Crotalid Venoms / Q000737:chemistry*; D010450:Endopeptidases / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008297:Male; D008833:Microcirculation / Q000187:drug effects; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D016415:Sequence Alignment
chemical_list: D000806:Angiotensin-Converting Enzyme Inhibitors; D003435:Crotalid Venoms; D010455:Peptides; D011498:Protein Precursors; D010450:Endopeptidases; D001920:Bradykinin