Structural studies of .alpha.-bungarotoxin. 3. Corrections in the primary sequence and x-ray structure and characterization of an isotoxic .alpha.-bungarotoxin
- 19 April 1988
- journal article
- research article
- Published by American Chemical Society (ACS) in Biochemistry
- Vol. 27 (8), 2775-2781
- https://doi.org/10.1021/bi00408a018
Abstract
The most plausible set of chemical shift assignments for .alpha.-bungarotoxin as deduced from the combined use of two-dimensional J-correlated and two-dimensional nuclear Overhauser effect 1H nuclear magnetic resonance (NMR) spectroscopy was in conflict with the accepted amino acid sequence between residues 8 and 12 and residues 66 and 70 [Basus, V. J., Billeter, M., Love, R. A., Stroud, R. M., and Kuntz, I. D. (1988) Biochemistry (first paper of three in this issue)]. Furthermore, NMR spectra of .alpha.-bungarotoxin, purified by conventional methods, evidenced a second species at the level of approximately 10% total protein. The minor component was separated from .alpha.-bungarotoxin by Mono-S (cationic) chromatography. Sequencing of Mono-S-purified .alpha.-bungarotoxin and one of its tryptic peptides showed that the correct sequence for .alpha.-bungarotoxin is Ser-Pro-Ile at positions 9-11 and Pro-His-Pro at positions 67-69. The electron density map of .alpha.-bungarotoxin [Love, R. A., and Stroud, R. M., (1986) Protein Eng. 1, 37] was refined with the new sequence data. Improvements in the structure were found primarily for residues 9-11. Sequence analysis of two overlapping tryptic peptides proved that the minor species differed from .alpha.-bungarotoxin by replacement of a valine for an alanine at position 31. This new toxin, .alpha.-bungarotoxin(Val-31), binds to the acetylcholine receptor with an affinity that is comparable to that of .alpha.-bungarotoxin.This publication has 17 references indexed in Scilit:
- Assignment of the 1H nuclear magnetic resonance spectrum of the proteinase inhibitor IIA from bull seminal plasma by two-dimensional nuclear magnetic resonance at 500 MHzJournal of Molecular Biology, 1983
- SECONDARY STRUCTURE AND SEQUENTIAL RESONANCE ASSIGNMENTS IN TWO-DIMENSIONAL PROTON MAGNETIC-RESONANCE SPECTRA OF INSECTOTOXIN-I5A BUTHUS-EUPEUS1983
- Blockade of transmission in rat sympathetic ganglia by a toxin which co-purifies with α-bungarotoxinBrain Research, 1982
- Individual Assignments of the Amide Proton Resonances Involved in the Triple‐Stranded Antiparallel Pleated β‐Sheet Structure of a Long Neurotoxin, Laticauda Semifasciata I11 from Laticauda semifasciataEuropean Journal of Biochemistry, 1982
- Sequential resonance assignments in protein 1H nuclear magnetic resonance spectraJournal of Molecular Biology, 1982
- Sequential resonance assignments in protein 1H nuclear magnetic resonance spectraJournal of Molecular Biology, 1982
- Proton‐Nuclear‐Magnetic‐Resonance Study on Molecular Conformations of Long NeurotoxinsEuropean Journal of Biochemistry, 1981
- Three-dimensional structure of the "long" neurotoxin from cobra venom.Proceedings of the National Academy of Sciences, 1980
- Inhibition of neuronal acetylcholine sensitivity by α-toxins from Bungarus multicinctus venomProceedings of the National Academy of Sciences, 1979
- Neurotoxins of Bungarus multicinctus venom purification and partial characterizationBiochemistry, 1977