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 μg·μL−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 μg·μL−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.

Peptidomics of Three Bothrops Snake Venoms: Insights Into the Molecular Diversification of Proteomes and Peptidomes

“…In light of previous reports on the structure and function relationship of BPPs (50,60,90,94), our current data show that the presence of a pyroglutamic acid residue at the N-terminal position and a Pro-Pro doublet at the C terminus in BPP sequences are not enough requirements for the peptide to display BK potentiating activity.…”

ESI-MS/MS Identification of a Bradykinin-Potentiating Peptide from Amazon Bothrops atrox Snake Venom Using a Hybrid Qq-oaTOF Mass Spectrometer

“…The measured peptide monoisotopic mass (1384.7386) and theoretical (1384.7378) was very similar, showing a mass accuracy of 0.6 ppm, which was also observed for the identified diagnostic fragment ions (Table 1, Table 2), thus showing the high precision of the analysis. Sequence similarity showed that the peptide is homologous to other BPP described for B. moojeni venom [6] and similar to others from Bothrops neuwiedi [1,9], B. leucurus , B. erythromelas , B. alternatus [10], B. insularis [1,10,11], B. jararaca [12,13], B. jararacussu [1,10,14], B. cotiara [13], and B. fonsecai [13] (Table 3). This peptide was named as Bradykinin-potentiating peptide BAX12.…”

Metallopeptidase inhibition potentiates bradykinin-induced hyperalgesia

“…Later studies confirmed that EP24.15 and EP24.16 are present in the cerebral vasculature and are capable of metabolizing BK in vivo [25,26]. Similarly, other peptidergic inhibitors of EP24.15 have demonstrated significant vasodilation effects when used in conjunction with BK, over using BK alone [29]. These findings are supportive of the work presented here, illustrating that EP24.15 and EP24.16 serve as important negative modulators of B2R activation by BK.…”