Cloning and sequence analysis of a
            <i>Bothrops jararaca</i>
            cDNA encoding a precursor of seven bradykinin-potentiating peptides and a C-type natriuretic peptide

Murayama, Nobuhiro; Hayashi, Mirian A. F.; Ohi, Hiroshi; Ferreira, L.A.F.; Hermann, V.V; Saito, Hiromi; Fujita, Yoshiaki; Higuchi, Shigesada; Fernandes, Beatriz Lieblich; Yamane, Toshimi; Camargo, Antônio Carlos Martins de

doi:10.1073/pnas.94.4.1189

Cited by 154 publications

(104 citation statements)

References 30 publications

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“…Moreover, other LAAO fragments have been found in the venoms of BC, BF and BJ (Table I), similarly as previously described in the venoms of B. moojeni (31,57) and B. insularis (58). All other peptides found by de novo sequencing were homologous or identical to sequences present in the BPP-CNP precursor which is composed of seven BPP molecules aligned in tandem after a hydrophobic signal peptide sequence, followed by a putative intervening sequence and a C-type natriuretic peptide at the C terminus (59). The BPP spectra are characterized by the typical m/z fragment at 213.1, corresponding to the y 2 of the C-terminal PP (8,20,31).…”

Section: De Novo Sequencing Of Peptides Present In the Venomssupporting

confidence: 55%

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

Tashima

Zelanis

Kitano

et al. 2012

Molecular & Cellular Proteomics

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Snake venom proteomes/peptidomes are highly complex and maintenance of their integrity within the gland lumen is crucial for the expression of toxin activities. There has been considerable progress in the field of venom proteomics, however, peptidomics does not progress as fast, because of the lack of comprehensive venom sequence databases for analysis of MS data. Therefore, in many cases venom peptides have to be sequenced manually by MS/MS analysis or Edman degradation. This is critical for rare snake species, as is the case of Bothrops cotiara (BC) and B. fonsecai (BF), which are regarded as near threatened with extinction. In this study we conducted a comprehensive analysis of the venom peptidomes of BC, BF, and B. jararaca (BJ) using a combination of solid-phase extraction and reversed-phase HPLC to fractionate the peptides, followed by nano-liquid chromatography-tandem MS (LC-MS/MS) or direct infusion electrospray ionization-(ESI)-MS/MS or MALDI-MS/MS analyses. We detected marked differences in the venom peptidomes and identified peptides ranging from 7 to 39 residues in length by de novo sequencing. Forty-four unique sequences were manually identified, out of which 30 are new peptides, including 17 bradykinin-potentiating peptides, three poly-histidine-poly-glycine peptides and interestingly, 10 L-amino acid oxidase fragments. Some of the new bradykinin-potentiating peptides display significant bradykinin potentiating activity. Automated database search revealed fragments from several toxins in the peptidomes, mainly from L-amino acid oxidase, and allowed the determination of the peptide bond specificity of proteinases and amino acid occurrences for the P4-P4 sites. We also demonstrate that the venom lyophilization/resolubilization process greatly increases the complexity of the peptidome because of the imbalance caused to the venom proteome and the consequent activity of proteinases on venom components. The use of proteinase inhibitors clearly showed different outcomes in the peptidome characterization and suggested that degradomic-peptidomic analysis of snake venoms is highly sensitive to the conditions of sampling procedures. Molecular & Cellular Proteomics 11: 10.1074/mcp.M112.019331, 1245-1262, 2012.Snake venoms are the products of specialized secretory glands located above the upper jawbone in venomous snakes. Like most secretory proteins, venom toxins are synthesized in the cytoplasm of secretory cells in the gland and transferred to the rough endoplasmic reticulum, then to the Golgi apparatus, and finally transported via secretory granules to the lumen of the venom gland (1). The snake venomous secretion is an aqueous solution containing a high amount of proteins and peptides, however the mechanisms for controlling protein secretion into the gland lumen and for regulating its protein/peptide concentration, ionic strength, and pH are unknown. As in all eukaryotic cells the proteomes of the venom gland tissue are highly complex, comprising a much great number and diversity of multidomain proteins (2...

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Section: De Novo Sequencing Of Peptides Present In the Venomssupporting

confidence: 55%

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

Tashima

Zelanis

Kitano

et al. 2012

Molecular & Cellular Proteomics

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“…This molecule, named Bj- or BPP-10c, belongs to a class of peptides isolated from Bothrops jararaca snake venom. These peptides are products of C-type natriuretic peptide precursor protein cleavage in the venom gland and the brain of the snake 8,9 Bj-PRO-10c exerted powerful and sustained antihypertensive activity 10 and vasodilatation depended on the positive modulation of ASS activity and NO production in the endothelium. 11 Changes in mean arterial pressure (MAP) promoted by Bj-PRO-10c are accompanied by a significant reduction in heart rate (HR), 10 rather than by an increase in HR as would be expected by the discharge of baroreceptors due to hypotension.…”

Section: Introductionmentioning

confidence: 99%

Brain nitric oxide production by a proline-rich decapeptide from Bothrops jararaca venom improves baroreflex sensitivity of spontaneously hypertensive rats

Lameu

Pontieri

Guerreiro³

et al. 2010

Hypertens Res

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Baroreflex sensitivity is disturbed in many people with cardiovascular diseases such as hypertension. Brain deficiency of nitric oxide (NO), which is synthesized by NO synthase (NOS) in the citrulline-NO cycle (with argininosuccinate synthase (ASS) activity being the rate-limiting step), contributes to impaired baroreflex. We recently showed that a decapeptide isolated from Bothrops jararaca snake venom, denoted Bj-PRO-10c, exerts powerful and sustained antihypertensive activity. Bj-PRO-10c promoted vasodilatation dependent on the positive modulation of ASS activity and NO production in the endothelium, and also acted on the central nervous system, inducing the release of GABA and glutamate, two important neurotransmitters in the regulation of autonomic systems. We evaluated baroreflex function using the regression line obtained by the best-fit points of measured heart rate (HR) and mean arterial pressure (MAP) data from spontaneously hypertensive rats (SHRs) treated with Bj-PRO-10c. We also investigated molecular mechanisms involved in this effect, both in vitro and in vivo. Bj-PRO-10c mediated an increase in baroreflex sensitivity and a decrease in MAP and HR. The effects exerted by the peptide include an increase in the gene expression of endothelial NOS and ASS. Bj-PRO-10c-induced NO production depended on intracellular calcium fluxes and the activation of a G i/o -protein-coupled metabotropic receptor. Bj-PRO-10c induced NO production and the gene expression of ASS and endothelial NOS in the brains of SHRs, thereby improving baroreflex sensitivity. Bj-PRO-10c may reveal novel approaches for treating diseases with impaired baroreflex function. Hypertension Research (2010Research ( ) 33, 1283Research ( -1288 doi:10.1038/hr.2010 Keywords: argininosuccinate synthase; baroreflex sensitivity; Bothrops jararaca venom; nitric oxide; proline-rich oligopeptide INTRODUCTIONThe gaseous messenger NO is involved mainly in the regulation of local and systemic vascular resistance, sodium balance and, consequently, blood pressure control, 1 but it is also a signaling molecule and modulator of brain function. 2 Recent studies relate NO with central cardiovascular control through the regulation of the cardiac and vascular autonomic system via the modulation of the central sites of cardiovascular autonomic neural integration. 3,4 NO is generated in the citrulline-NO cycle by NO synthase (NOS) using L-arginine as a substrate. Three isoforms of NOS have been described: calcium-dependent endothelial (eNOS) and neuronal (nNOS) isoforms and inducible NOS. The expression and activity

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“…Three reasons explain the renewed interest: a number of the Bj-BPPs can distinguish between the N and C active sites of sACE (Cotton et al, 2002). In fact, sACE has two homologous and functionally distinct active sites (Wei et al, 1991), one of which, the active site at the C domain (C site), is slightly more effective in hydrolyzing some vasoactive peptides, like bradykinin and angiotensin I (Perich et al, 1992;Jaspard et al, 1993); 2) the isolation and identification of novel Bj-BPPs in the crude venom (Ianzer et al, 2004); and 3) the presence of several Bj-BPPs in the C-type natriuretic peptide precursor of the snake brain may reveal novel neuropeptides (Murayama et al, 1997;Hayashi et al, 2003).…”

mentioning

confidence: 99%

“…We have previously described the isolation of a cDNA coding for a Bj-BPP precursor protein from the venom gland and found in the neuroendocrine areas of the Bj-brain, containing seven tandemly arranged Bj-BPP sequences at the N terminus and one C-type natriuretic peptide sequence at the C terminus (C site) (Murayama et al, 1997;Hayashi et al, 2003). Among the endogenous, BPP-9a and BPP-10c turned out to be the most efficient inhibitors of the sACE, displaying high selectivity toward the C-active site (Cotton et al, 2002;Hayashi et al, 2003).…”

mentioning

confidence: 99%

Do the Cardiovascular Effects of Angiotensin-Converting Enzyme (ACE) I Involve ACE-Independent Mechanisms? New Insights from Proline-Rich Peptides ofBothrops jararaca

Ianzer

Santos²,

Etelvino³

et al. 2007

J Pharmacol Exp Ther

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Angiotensin-converting enzyme (ACE) inhibitors were developed based on proline-rich oligopeptides found in the venom of Bothrops jararaca (Bj) previously known as bradykinin-potentiating peptides (BPPs). However, the exact mechanism of action of BPPs remains unclear. The role of the ACE in the cardiovascular effects of two of naturally proline-rich oligopeptides (Bj-BPP-7a and Bj-BPP-10c) was evaluated in vitro and in vivo. Bj-BPP-7a does not potentiate the cardiovascular response to bradykinin and is a weak inhibitor of ACE C and N sites (K i ϭ 40,000 and 70,000 nM, respectively), whereas Bj-BPP-10c is a strong bradykinin potentiator and inhibitor of the ACE C site (K i ϭ 0.5 versus 200 nM for N site). Strikingly, both peptides, in doses ranging from 0.47 to 71 nmol/kg, produced long-lasting reduction (Ͼ6 h) in the mean arterial pressure of conscious spontaneously hypertensive rats (maximal change, 45 Ϯ 6 and 53 Ϯ 6 mm Hg for Bj-BPP-7a and Bj-BPP-10c, respectively). The fall in blood pressure was accompanied by variable degrees of bradycardia. In keeping with the absence of relationship between ACE-inhibitory and antihypertensive activities, no changes in the pressor effect of angiotensin I or in the hypotensive effect of bradykinin were observed at the peak of the cardiovascular effects of both peptides. Our results indicate that the antihypertensive effect of two Bj-BPPs containing the motif Ile-Pro-Pro is unrelated to their ability for inhibiting ACE or potentiating bradykinin (BK), indicating as a major component ACE and BK-independent mechanisms. These results are in line with previous observations suggesting ACE inhibition-independent mechanisms for angiotensin I-converting enzyme inhibitor.

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Cloning and sequence analysis of a Bothrops jararaca cDNA encoding a precursor of seven bradykinin-potentiating peptides and a C-type natriuretic peptide

Cited by 154 publications

References 30 publications

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

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

Brain nitric oxide production by a proline-rich decapeptide from Bothrops jararaca venom improves baroreflex sensitivity of spontaneously hypertensive rats

Do the Cardiovascular Effects of Angiotensin-Converting Enzyme (ACE) I Involve ACE-Independent Mechanisms? New Insights from Proline-Rich Peptides ofBothrops jararaca

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