Biochemical and Biophysical Research Communications
Identification and functional analysis of a novel bradykinin inhibitory peptide in the venoms of New World Crotalinae pit vipers
Section snippets
Materials and methods
Reagents. All solvents were of HPLC grade unless otherwise stated and purchased from Sigma (Poole, Dorset, UK) except acetic acid, which was obtained from BDH (Poole, Dorset, UK).
Acquisition of venoms. All venoms were collected from authenticated snake species housed at the Alistair Reid Venom Unit of the Liverpool School of Tropical Medicine, University of Liverpool (Liverpool, UK), and were supplied in a lyophilized form.
Gel permeation chromatography. Lyophilized snake venoms (9–10 mg dry
Chromatographic fractionation of snake venoms
Thirty snake venoms were fractionated using Sephadex G50 gel permeation chromatography (data not shown). Three of these venoms, from A. bilineatus (Mexican moccasin), C. v. viridis (prairie rattlesnake), and L. muta (bushmaster), displayed striking similarities in their gel permeation profiles (Fig. 1). These all displayed a penultimate peak of absorbance that was larger than the final, total volume peak of absorbance (Fig. 1). Pooled fractions representing this penultimate peak from the three
Discussion
In this study, the low molecular weight peptides present in gel permeation chromatographic fractions of the venoms from three species of New World pit vipers from the subfamily, Crotalinae, that is the Mexican moccasin (A. bilineatus), the prairie rattlesnake (C. v. viridis), and the South American bushmaster (L. muta), were characterized using HPLC, MALDI-TOF mass spectrometry, and Edman sequencing. This strategy led to the identification of a novel peptide entity that we named bradykinin
Acknowledgment
We thank the Department of Education Northern Ireland for funding the PhD studentship of Robert Leslie Graham.
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Unveiling the peptidome diversity of Lachesis muta snake venom: Discovery of novel fragments of metalloproteinase, L-amino acid oxidase, and bradykinin potentiating peptides
2023, Biochemical and Biophysical Research CommunicationsHalf a century of research on Bothrops asper venom variation: biological and biomedical implications
2023, ToxiconCitation Excerpt :BPPs are associated with the hypotensive effect of some botropic envenomings because they act mainly at the level of the renin-angiotensin and kallikrein-kinin systems by inhibiting angiotensin-I converting enzyme (ACE), which in turn leads to vasodilation events, increased capillary permeability, and hypovolemic shock (Ferreira et al., 1970; Greene et al., 1972; Dendorfer et al., 2000; Luft, 2008; Koh and Kini, 2012; Munawar et al., 2018). On the other hand, SVMPi are endogenous tripeptides derived proteolytically from the N-terminus of certain BPPs (James Graham et al., 2005; Cidade et al., 2006), that act as inhibitors of the metalloprotease activity of the venom at the glandular level, whose effectiveness is deactivated by dilution or physicochemical change when the venom is injected into the prey or the victim (Huang et al., 1998, 2002; Munekiyo and Mackessy, 2005; Wagstaff et al., 2008). Tripeptides pENW and pEQW, similar to those of B. jararaca, and with the ability to inhibit the protease activity of the same venom, have been isolated from B. asper venom (Francis and Kaiser, 1993) or identified by mass spectrometry (Mora-Obando et al., 2020).
Venomics of the poorly studied hognosed pitvipers Porthidium arcosae and Porthidium volcanicum
2021, Journal of ProteomicsCitation Excerpt :In addition, both proteomes contain disparate amount of endogenous tripeptide inhitors of SVMPs (SVMPi, 0.6% in P.arcosae but 6.3% in P. volcanicum) and bradykinin-potentiating and inhibitory peptides (BPP (0.8%) and BIP (0.7%), respectively) were also identified in P. arcosae but not in P. volcanicum venom. SVMPi, BIP and BPPs are released into the venom proteome by the proteolytic processing of a common precursor [72,73]. SVMPi reversibly bind to and inactivate SVMPs stored at high concentration in the lumen of the venom gland of many Viperidae snakes, protecting glandular tissues and venom factors from the proteolysis [72–77].