Summary
A nonapeptide Thr-Ile-Ile-Asn-Val-Lys-Cys-Thr-Ser (NTX1–9) and a decapeptide Met-Asn-Gly-Lys-Cys-Lys-Cys-Tyr-Asn-Asn (NTX30–39) corresponding to the N-terminal and C-terminal sequences respectively of Noxiustoxin (NTX) were synthesized by the solid phase method of Merrifield (1963). The first synthetic peptide (NTX1–9) was shown to be toxic to mice independently of the route of administration: intraperitoneally, subcutaneously or intraventricularly (100–200 μg/20 g mouse weight). The second (NTX30–39) was not toxic even at higher dose (400 μg/20 g mouse). When the effects of the peptide NTX1–9 and of the authentic toxin (Noxiustoxin) were studied on the liberation of [3H] 4-aminobutyric acid (3H-GABA) from mouse synaptosomes, both gave essentially the same results, except that peptide NTX1–9 was needed at higher concentration. Synthetic peptide NTX30–39 had no effect in the same preparation at even higher doses. The GABA release produced by toxic peptide NTX1–9 was not affected by tetrodotoxin but was completely abolished by the presence of the K+ ionophore valinomycin, mimicking the effect of native NTX in the same system (Sitges et al., 1986). These results indicate that the toxic active site of Noxiustoxin is possibly located in or near the N-terminal amino acid portion of the molecule.
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Abbreviations
- BOC:
-
amino acids ter-butyloxycarbonyl-amino acids
- BOC:
-
amino acid-PAM-resin ter-butyloxycarbonyl-aminoacyl-4-(oxymethyl)-phenacetamidomethyl-resin
- GABA 4:
-
aminobutyric acid
- HPLC:
-
high performance liquid chromatography
- MSA:
-
mouse serum albumin
- NTX:
-
Noxiustoxin
- NTX:
-
(numbers) synthetic peptides with amino acid sequences of NTX at position start (first number) to position end (second number) of the sequence according to Fig. 1
- TTX:
-
tetrodotoxin
References
Carbone E, Wanke E, Prestipino G, Possani LD, Maelicke A (1982) Selective blockage of voltage-dependent K+ channels by a novel scorpion toxin. Nature 296: 90–91
Carbone E, Prestipino G, Spadavecchia F, Franciolini F, Possani LD (1987) Blocking of the squid axon K+ channel by Noxiustoxin: a toxin from the venom of the scorpionCentruroides noxius. Pfiuegers Arch (Eur J Physiol) 408: 423–431
Edman P, Begg G (1967) A protein sequenator. Eur J Biochem 1: 80–91
Esko K, Karlsson M (1970) Effects of Boc-amino acid adsorption with respect to yield and racemization in the Merrifield method. Acta Chem Scand 24: 1415–1422
Gurrola B, Herion P, Possani LD (1988) Manuscript in preparation
Hajós F (1975) An improved method for the preparation of synaptosomal fractions in high purity. Brain Res 93: 485–489
Merrifield BR (1963) Solid phase peptide synthesis I. The synthesis of a tetrapeptide. J Am Chem Soc 85: 2144–2154
Mitchell AR, Kent SBH, Engelhard M, Merrifield BR (1978) A new synthetic rute to terbutyloxycarbonylaminoacyl-4-(oxymethyl)phenylacetamidomethyl-resin and improved support for solid phase peptide synthesis. J Org Chem 43: 2845–2852
Moore A, Stein WH (1963) Chromatographic determination of amino acids by the use of automatic recording equipment In: Colowick SP, Kaplan NO (eds) Methods in enzymology, vol 6. Academic Press, New York, pp 819–831
Possani LD, Dent MAR, Martin BM, Maelicke A, Svendsen I (1981) The amino terminal sequence of several toxins from the venom of the Mexican scorpionCentruroides noxius Hoffmann. Carlsberg Res Commun 46: 207–214
Possani LD, Martin BM, Svendsen I (1982) The primary structure of Noxiustoxin: a K+ channel blocker peptide, purified from the venom of the scorpionCentruroides noxius Hoffmann. Carlsberg Res Commun 47: 285–289
Possani LD, Martin BM, Svendsen I, Rode GS, Erickson BW (1985) Scorpion toxins fromCentruroides noxius andTityus serrulatus. Biochem J 229: 739–750
Robinson AB (1967) Ph.D. thesis presented to University of California, San Diego, CA, U.S.A.
Sarin VK, Kent SBH, Tam JP, Merrifield BR (1981) Quantitative monitoring of solid-phase peptide synthesis by ninhydrin reaction. Anal Biochem 117: 147–157
Sitges M, Possani LD, Bayon A (1986) Noxiustoxin, a short-chain toxin from the Mexican scorpionCentruroides noxius, induces transmitter release by blocking K+ permeability. J Neurosci 6: 1570–1574
Valdivia HH, Martin BM, Possani LD (1988) Labelling of Noxiustoxin, a K channel blocker peptide, and its binding properties to mouse brain nerve-ending particles. Neurochemistry (submitted for publication)
Valdivia HH, Smith JS, Martin BM, Coronado R, Possani LD (1988) Charybdotoxin and Noxiustoxin, two homologous peptide inhibitors of the K+ (Ca++) channel. FEBS Lett 226: 280–284
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Supported in part by the Mexican Council of Science and Technology (CONACyT), grants PVT/QF/NAL/84/2182, PVT/AI/NAL/85/3029 to L.D.P.; PCSACNA 022640 to A.B. A patent request claiming rights on the use of synthetic NTX and related peptides was presented in Washington, DC (U.S.A.), Serial number 07/132,169, filing date December 14, 1987.
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Gurrola, G.B., Molinar-Rode, R., Sitges, M. et al. Synthetic peptides corresponding to the sequence of noxiustoxin indicate that the active site of this K+ channel blocker is located on its amino-terminal portion. J. Neural Transmission 77, 11–20 (1989). https://doi.org/10.1007/BF01255815
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DOI: https://doi.org/10.1007/BF01255815