Abstract
Type A botulinum neurotoxin catalyzed the hydrolysis of synthetic peptides based on the sequence of the 25-kD synaptosomal protein SNAP-25. In each peptide, the toxin cleaved at a single glutaminyl-arginine bond corresponding to residues 197 and 198 of SNAP-25, confirming earlier reports on the enzymatic specificity of the toxin in synaptosomal preparations. Metal chelators inhibited catalysis, consistent with a metalloprotease activity. In contrast to tetanus toxin and other botulinum toxin serotypes, type A toxin hydrolyzed relatively short, 17-to 20-residue peptides. In the substrates, SNAP-25 residue 202 and one or more of residues 187–191 were required for efficient hydrolysis, but residues 167–186 and 203–206 were not. The highest rates of hydrolysis were found when the C-terminal residues of the peptides were amidated.
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Blasi, J., Chapman, E. R., Link, E., Binz, T., Yamasaki, S., De Camilli, P., Sudhof, T. C., Nieman, H., and Jahn, R. (1993).Nature 365, 160–163.
Cornille, F., Goudreau, N., Ficheux, D., Nieman, H., and Roques, B. (1994).Eur. J. Biochem. 222, 173–181.
Dolly, J. O. (1992). In:Handbook of Experimental Pharmacology (Herken, H., and Hucho, F., eds.), Springer-Verlag, Berlin, pp. 681–717.
Eipper, B. A., and Mains, R. E. (1988).Annu. Rev. Physiol. 50, 333–344.
Eisel, U., Jarausch, W., Goretzki, K., Henschen, A., Engels, J., Weller, U., Hudel, M., Habermann, E., and Nieman, H. (1986).EMBO J. 5, 2495–2502.
Foran, P., Shone, C. C., and Dolly, J. O. (1994).Biochemistry 33, 15365–15374.
Hansen, S., Kristian, L., and Hough, E. (1993).J. Mol. Biol. 231, 870–876.
Jankovic, J., and Brin, M. F. (1992).N. Engt J. Med. 324, 1186–1194.
Jongeneel, C. V., Bouvier, J., and Bairoch, A. (1989).FEBS Lett. 242, 211–214.
Nieman, H. (1991). InSourcebook of Bacterial Protein Toxins (Alouf, J., and Freer, J., eds.), Academic Press, New York, pp. 303–348.
Nieman, H., Blasi, J., and Jahn, R. (1994).Trends Cell Biol. 4, 179–185.
Oyler, G. A., Higgins, G. A., Hart, R. A., Battenberg, E., Billingsley, M., Bloom, F. E., and Wilson, M. C. (1989).J. Cell Biol. 109, 3039–3052.
Rosetto, O., Schiavo, G., Montecucco, C., Poulain, B., Deloye, F., Lozzi, L., and Shone, C. C. (1994).Nature 372, 415–416.
Schiavo, G., Benfenatti, F., Poulain, B., Rossetto, O., deLaureto, P. P., DasGupta, B. R., and Montecucco, C. (1992a).Nature 359, 832–835.
Schiavo, G., Poulain, B., Rossetto, O., Benfenatti, F., Tauc, L., and Montecucco, C. (1992b).EMBO J. 11, 3577–3583.
Schiavo, G., Rossetto, O., Catsicas, S., de Laureto, P. P., DasGupta, B. R., Benfenati, F., and Montecucco, C. (1993).J. Biol. Chem. 268, 23784–23787.
Schiavo, G., Rossetto, O., and Montecucco, C. (1994).Cell Biol. 5, 221–229.
Shone, C. C., and Roberts, A. K. (1994).Eur. J. Biochem. 225, 263–270.
Shone, C. C., Quinn, C. P., Wait, R., Hallis, B., Fooks, S., and Hambleton, P. (1993).Eur. J. Biochem. 217, 965–971.
Simpson, L. L. (1981).Pharmacol. Rev. 33, 155–188.
Simpson, L. L. (1984).J. Pharmacol. Exp. Ther. 230, 665–669.
Yamasaki, S., Baumeister, A., Binz, T., Blasi, J., Link, E., Cornille, F., Rocques, B., Fykse, E. M., Sudhof, T. C., Jahn, R., and Nieman, H. (1994).J. Biol. Chem. 269, 12764–12772.
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Schmidt, J.J., Bostian, K.A. Proteolysis of synthetic peptides by type A botulinum neurotoxin. J Protein Chem 14, 703–708 (1995). https://doi.org/10.1007/BF01886909
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DOI: https://doi.org/10.1007/BF01886909