Immunological and structural characterization of sarafotoxin/endothelin family of peptides

Fleminger, Gideon; Bousso-Mittler, D.; Bdolah, Avner; Kloog, Yoel; Sokolovsky, Mordechai

doi:10.1016/0006-291x(89)90817-6

Cited by 15 publications

(6 citation statements)

References 12 publications

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“…113,114 The chemical structure of the endothelins is closely related to neurotoxins (sarafotoxins) produced by scorpions and snakes. [115][116][117] Factors modulating the expression of ET-1 are shearstress, adrenaline, angiotensin II, thrombin, inflammatory cytokines (tumor necrosis factor-␣, interleukin-1 and -2), transforming growth factor-␤, and hypoxia. [118][119][120][121][122][123][124][125][126][127][128][129][130] ET-1 is metabolized by a neutral endopeptidase, which also cleaves natriuretic peptides.…”

Section: Endothelinmentioning

confidence: 99%

Working under pressure: the vascular endothelium in arterial hypertension

et al. 2000

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The vascular endothelium synthesizes and releases a spectrum of vasoactive substances like nitric oxide (NO) and endothelin (ET). In hypertension, the delicate balance of endothelium-derived factors is disturbed. ET acts as the natural counterpart to endothelium-derived NO, which exerts vasodilating, antithrombotic, and antiproliferative effects, and inhibits leukocyte-adhesion to the vascular wall. Besides its blood pressure rising effect also in man, ET induces vascular and myocardial hypertrophy, which are independent risk factors for cardiovascular morbidity and mortality. The derangement of endothelial function in hypertension is likely to be caused in part by genetic factors, but also due to elevated blood pressure itself. Due to its position between blood pressure and smooth muscle cells responsible for peripheral resistance, the endothelium is thought to be both target and mediator of arterial hypertension. Oxi-

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Section: Endothelinmentioning

confidence: 99%

Working under pressure: the vascular endothelium in arterial hypertension

et al. 2000

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“…Analysis of peptides ,g in 0.1 ml of0. 1% trifluoroacetic acid (TFA) in water] was performed as described (21) …”

Section: Methodsmentioning

confidence: 99%

“…The endothelins (ETs) and the sarafotoxins (SRTXs) are two classes of a recently discovered family of peptides containing 21 amino acids (reviewed in refs. 1 and 2).…”

mentioning

confidence: 99%

“…These considerations prompted us to examine whether the ETs and/or the SRTXs might be good substrates for NEP. The effect of the enzyme can be evaluated by measuring enzyme-associated changes in the biochemical activities of the peptides, such as binding to the receptor, induction of inositol phospholipid hydrolysis, immunogenicity (20,21), and lethality. As shown in this in vitro study, NEP inactivates ETs as a result of nicking the bond at Ser5-Leu6 followed by rapid cleavage at the amino side of Ile"9.…”

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confidence: 99%

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Endothelins are more sensitive than sarafotoxins to neutral endopeptidase: possible physiological significance.

Skolovsky

Galron

Kloog

et al. 1990

Proc. Natl. Acad. Sci. U.S.A.

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Incubation of endothelins (ETs) with bovine kidney neutral endopeptidase (NEP) resulted in a selective two-step degradation with loss of biochemical activity. The Km of the enzyme indicated high-affinity binding, and hydrolysis was completely inhibited by phosphoramidon. The first step was nicking of the Ser5-Leu' bond, followed by cleavage at the amino side of ile". The nicked peptide exhibited biochemical activities comparable to those of the intact peptide-i.e., binding to the ET receptor, induction of inositol phospholipid hydrolysis, and toxicity. The twice-cleaved product was inactive. The sarafotoxins (SRTXs) were more resistant than the ETs to NEP: for example, the half-time for ET-1 was 41 hr, while it was =4 hr for SRTX-b and even higher for SRTX-c. These in vitro rmdings may indicate a regulatory role of NEP (or similar enzymes) in the physiological inactivation of ETs. They might also help to explain why under certain physiological conditions ETs may be less toxic than SRTXs.The endothelins (ETs) and the sarafotoxins (SRTXs) are two classes of a recently discovered family of peptides containing 21 amino acids (reviewed in refs. 1 and 2). The ETs (isolated from mammalian systems) and the SRTXs (from snake venom) are among the most potent vasoconstrictor agents known (refs. 1 and 2 and references therein) and are highly lethal (3). The two classes of peptides show a considerable sequence homology: they each possess four cysteinyl residues, and "'60%o of the 21 amino acid residues are common to both. Since ET-1 was described, three other ETs have been detected and are designated ET-2, ET-3 (4), and (Vic) (5) (Fig. 1). Also, four SRTXs-SRTX-a, SRTX-b, SRTX-c, and SRTX-d (6)-have been characterized so far (Fig. 1). The four cysteinyl residues in the ETs and in the SRTXs are interconnected by disulfide bridges between positions 1 and 15 and 3 and 11, forming an intramolecular loop structure (see diagram in Fig. 1 (19). The substrates cleaved by NEP appear to be restricted to peptides of molecular mass ==3 kDa (18,19). These considerations prompted us to examine whether the ETs and/or the SRTXs might be good substrates for NEP. The effect of the enzyme can be evaluated by measuring enzyme-associated changes in the biochemical activities of the peptides, such as binding to the receptor, induction of inositol phospholipid hydrolysis, immunogenicity (20, 21), and lethality. As shown in this in vitro study, NEP inactivates ETs as a result of nicking the bond at Ser5-Leu6 followed by rapid cleavage at the amino side of Ile"9. Under identical experimental conditions, NEP also inactivated SRTXs, but at a much slower rate. MATERIALS AND METHODSET-1, ET-2, ET-3, and ET-4 (Vic) were purchased from American Peptide (Santa Clara, CA). SRTX-b and SRTX-c were those used in a previous study (20). 125I-labeled ET-1 (125I-ET-1) was prepared by iodination with Enzymobeads (Bio-Rad) and was purified as described (8). Bovine kidney NEP (0.4 mg/ml) was purified by Triton X-100 extraction followed by DEAE-Sepharose Fast...

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“…Specifically, it is noted that the disulphide-reduced ET [29], fully Cysprotected ET [30] and [Ala ~, Ala a, Ala j J, Ala Is] ET [31]~ as well as the ET derivative formed by cleaving at Lys 9 with lysyl endopeptidase [29], all have greatly reduced potency by comparison with native ET-1. (It is surprising to observe, however, that there is no apparent loss in receptor binding For ET cleaved at Met 7 by cyanogen bromide treatment [33], but the full effects of this modification remain to be determined. )…”

Section: Discussion • and Conclusionmentioning

confidence: 99%

Molecular modelling of the structures of endothelin antagonists Identification of a possible structural determinant for ET‐A receptor binding

Satoh

Barlow

1992

FEBS Letters

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Computer-aided molecular modelling of the endothclin (ET-A) receptor antagonists, BQ-123 and BE-18257B. shows that they have very similar 3D structures. Parts of their 3D structures are also shown to match closely with that reported for residues 6-8 in endothelind. On the basis of these similarities (and with supporting evidence from literature data on endothelin structure-activity relationships) a structural determinant is proposed for ET-A receptor binding, and novel designs of peptide are suggested for providing more potent and selective ET-A receptor antagonists.

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Immunological and structural characterization of sarafotoxin/endothelin family of peptides

Cited by 15 publications

References 12 publications

Working under pressure: the vascular endothelium in arterial hypertension

Working under pressure: the vascular endothelium in arterial hypertension

Endothelins are more sensitive than sarafotoxins to neutral endopeptidase: possible physiological significance.

Molecular modelling of the structures of endothelin antagonists Identification of a possible structural determinant for ET‐A receptor binding

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