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A novel bradykinin potentiating peptide isolated from Bothrops jararacussu venom using catallytically inactive oligopeptidase EP24.15

2008; Wiley; Volume: 275; Issue: 10 Linguagem: Inglês

10.1111/j.1742-4658.2008.06389.x

1742-4658

Vanessa Rioli, Benedito C. Prezoto, Katsuhiro Konno, Robson L. Melo, Clécio F. Klitzke, Emer S. Ferro, Mônica Ferreira‐Lopes, Antônio Carlos Martins de Camargo, Fernanda Calheta Vieira Portaro,

Biochemical and Structural Characterization

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.