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. 1978 Oct 1;175(1):281–288. doi: 10.1042/bj1750281

beta-Bungarotoxin. The binding of [3H]pyridoxylated beta-bungarotoxin to a high-molecular-weight protein receptor.

J MacDermot, R H Westgaard, E J Thompson
PMCID: PMC1186064  PMID: 736898

Abstract

beta-Bungarotoxin was labelled with pyridoxal 5'-phosphate (incorporating 3H). The kinetics of beta-bungarotoxin binding to several tissue subfragments of nervous tissue was studied. The dissociation constant of 3H-pyridoxylated beta-bungarotoxin in this reaction was 0.21-0.37 micron and that of unlabelled beta-bungarotoxin was 25 nM. Hill [(1910) J. Physiol. (London) 40, iv-vii] and Scatchard [(1949) Ann. N.Y. Acad. Sci. 51, 660-672] analyses demonstrated no co-operativity of binding and only a single class of receptor sites, consistent with a bimolecular association of beta-bungarotoxin and its receptor. The iodinated toxin was physiologically inactive. Toxin was bound in non-specific unsaturable fashion by glass and/or plastic. This low-affinity binding was corrected by addition of bovine serum albumin to a final concentration of 30 mg/ml. A soluble protein receptor of beta-bungarotoxin was isolated and the mol.wt. is approx. 200000.

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Selected References

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  1. Anker H. S. A solubilizable acrylamide gel for electrophoresis. FEBS Lett. 1970 Apr 16;7(3):293–293. doi: 10.1016/0014-5793(70)80185-5. [DOI] [PubMed] [Google Scholar]
  2. Berl S., Puszkin S., Nicklas W. J. Actomyosin-like protein in brain. Science. 1973 Feb 2;179(4072):441–446. doi: 10.1126/science.179.4072.441. [DOI] [PubMed] [Google Scholar]
  3. Blitz A. L., Fine R. E. Muscle-like contractile proteins and tubulin in synaptosomes. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4472–4476. doi: 10.1073/pnas.71.11.4472. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burridge K., Bray D. Purification and structural analysis of myosins from brain and other non-muscle tissues. J Mol Biol. 1975 Nov 25;99(1):1–14. doi: 10.1016/s0022-2836(75)80154-9. [DOI] [PubMed] [Google Scholar]
  5. CHANG C. C., LEE C. Y. ISOLATION OF NEUROTOXINS FROM THE VENOM OF BUNGARUS MULTICINCTUS AND THEIR MODES OF NEUROMUSCULAR BLOCKING ACTION. Arch Int Pharmacodyn Ther. 1963 Jul 1;144:241–257. [PubMed] [Google Scholar]
  6. Chang C. C., Chen T. F., Lee C. Y. Studies of the presynaptic effect of -bungarotoxin on neuromuscular transmission. J Pharmacol Exp Ther. 1973 Feb;184(2):339–345. [PubMed] [Google Scholar]
  7. Chang K. J., Jacobs S., Cuatrecasas P. Quantitative aspects of hormone-receptor interactions of high affinity. Effect of receptor concentration and measurement of dissociation constants of labeled and unlabeled hormones. Biochim Biophys Acta. 1975 Oct 6;406(2):294–303. doi: 10.1016/0005-2736(75)90011-5. [DOI] [PubMed] [Google Scholar]
  8. Chen L. B. Alteration in cell surface LETS protein during myogenesis. Cell. 1977 Mar;10(3):393–400. doi: 10.1016/0092-8674(77)90026-5. [DOI] [PubMed] [Google Scholar]
  9. Churchich J. E. Energy transfer in protein pyridoxamine-5-phosphate conjugates. Biochemistry. 1965 Jul;4(7):1405–1410. doi: 10.1021/bi00883a027. [DOI] [PubMed] [Google Scholar]
  10. Cooper D., Reich E. Neurotoxin from venom of the cobra, Naja naja siamensis. Purification and radioactive labeling. J Biol Chem. 1972 May 25;247(10):3008–3013. [PubMed] [Google Scholar]
  11. DIXON M. The determination of enzyme inhibitor constants. Biochem J. 1953 Aug;55(1):170–171. doi: 10.1042/bj0550170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fine R. E., Blitz A. L., Hitchcock S. E., Kaminer B. Tropomyosin in brain and growing neurones. Nat New Biol. 1973 Oct 10;245(145):182–186. doi: 10.1038/newbio245182a0. [DOI] [PubMed] [Google Scholar]
  13. Fine R. E., Bray D. Actin in growing nerve cells. Nat New Biol. 1971 Nov 24;234(47):115–118. doi: 10.1038/newbio234115a0. [DOI] [PubMed] [Google Scholar]
  14. Howard B. D. Effects of beta-bungarotoxin on mitochondrial respiration are caused by associated phospholipase A activity. Biochem Biophys Res Commun. 1975 Nov 3;67(1):58–65. doi: 10.1016/0006-291x(75)90282-x. [DOI] [PubMed] [Google Scholar]
  15. Howard B. D., Wu W. C. Evidence that beta-bungarotoxin acts at the exterior of nerve terminals. Brain Res. 1976 Feb 13;103(1):190–192. doi: 10.1016/0006-8993(76)90704-6. [DOI] [PubMed] [Google Scholar]
  16. Kelly R. B., Oberg S. G., Strong P. N., Wagner G. M. beta-Bungarotoxin, a phospholipase that stimulates transmitter release. Cold Spring Harb Symp Quant Biol. 1976;40:117–125. doi: 10.1101/sqb.1976.040.01.013. [DOI] [PubMed] [Google Scholar]
  17. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  18. MacDermot J., Westgaard R. H., Thompson E. J. beta-Bungarotoxin. Separation of two discrete proteins with different synaptic actions. Biochem J. 1978 Oct 1;175(1):271–279. doi: 10.1042/bj1750271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Marchalonis J. J. An enzymic method for the trace iodination of immunoglobulins and other proteins. Biochem J. 1969 Jun;113(2):299–305. doi: 10.1042/bj1130299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Oberg S. G., Kelly R. B. Saturable binding to cell membranes of the presynaptic neurotoxin, beta-bungarotoxin. Biochim Biophys Acta. 1976 May 21;433(3):662–673. doi: 10.1016/0005-2736(76)90289-3. [DOI] [PubMed] [Google Scholar]
  22. Pearse B. M. Coated vesicles from pig brain: purification and biochemical characterization. J Mol Biol. 1975 Sep 5;97(1):93–98. doi: 10.1016/s0022-2836(75)80024-6. [DOI] [PubMed] [Google Scholar]
  23. Sen I., Grantham P. A., Cooper J. R. Mechanism of action of beta-bungarotoxin on synaptosomal preparations. Proc Natl Acad Sci U S A. 1976 Aug;73(8):2664–2668. doi: 10.1073/pnas.73.8.2664. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Strong P. N., Goerke J., Oberg S. G., Kelly R. B. beta-Bungarotoxin, a pre-synaptic toxin with enzymatic activity. Proc Natl Acad Sci U S A. 1976 Jan;73(1):178–182. doi: 10.1073/pnas.73.1.178. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wang K., Ash J. F., Singer S. J. Filamin, a new high-molecular-weight protein found in smooth muscle and non-muscle cells. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4483–4486. doi: 10.1073/pnas.72.11.4483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  27. Wernicke J. F., Vanker A. D., Howard B. D. The mechanism of action of beta-bungarotoxin. J Neurochem. 1975 Oct;25(4):483–496. doi: 10.1111/j.1471-4159.1975.tb04354.x. [DOI] [PubMed] [Google Scholar]
  28. Whittaker V. P., Michaelson I. A., Kirkland R. J. The separation of synaptic vesicles from nerve-ending particles ('synaptosomes'). Biochem J. 1964 Feb;90(2):293–303. doi: 10.1042/bj0900293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Willingham M. C., Ostlund R. E., Pastan I. Myosin is a component of the cell surface of cultured cells. Proc Natl Acad Sci U S A. 1974 Oct;71(10):4144–4148. doi: 10.1073/pnas.71.10.4144. [DOI] [PMC free article] [PubMed] [Google Scholar]

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