Abstract
The interaction of botulinum neurotoxin serotypes A, B and E with membranes of different lipid compositions was examined by photolabelling with two photoreactive phosphatidylcholine analogues that monitor the polar region and the hydrophobic core of the lipid bilayer. At neutral pH the neurotoxins interacted both with the polar head groups and with fatty acid chains of phospholipids. At acidic pHs the neurotoxins underwent structural changes characterized by a more extensive interaction with lipids. Both the heavy and light chain subunits of the neurotoxins were involved in the process. The change in the nature and extent of toxin-lipid interaction occurred in the pH range 4-6 and was not influenced by the presence of polysialogangliosides. The present data are in agreement with the idea that botulinum neurotoxins enter into nerve cells from a low pH intracellular compartment.
Full text
PDF






Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Agui T., Syuto B., Oguma K., Iida H., Kubo S. Binding of Clostridium botulinum type C neurotoxin to rat brain synaptosomes. J Biochem. 1983 Aug;94(2):521–527. doi: 10.1093/oxfordjournals.jbchem.a134383. [DOI] [PubMed] [Google Scholar]
- Agui T., Syuto B., Oguma K., Iida H., Kubo S. The structural relation between the antigenic determinants to monoclonal antibodies and binding sites to rat brain synaptosomes and GT1b ganglioside in Clostridium botulinum type C neurotoxin. J Biochem. 1985 Jan;97(1):213–218. doi: 10.1093/oxfordjournals.jbchem.a135047. [DOI] [PubMed] [Google Scholar]
- Bigalke H., Müller H., Dreyer F. Botulinum A neurotoxin unlike tetanus toxin acts via a neuraminidase sensitive structure. Toxicon. 1986;24(11-12):1065–1074. doi: 10.1016/0041-0101(86)90133-9. [DOI] [PubMed] [Google Scholar]
- Bisson R., Montecucco C., Capaldi R. A. Interaction of membranous cytochrome b5 with arylazidophospholipids. FEBS Lett. 1979 Oct 15;106(2):317–320. doi: 10.1016/0014-5793(79)80523-2. [DOI] [PubMed] [Google Scholar]
- Bisson R., Montecucco C. Photolabelling of membrane proteins with photoactive phospholipids. Biochem J. 1981 Mar 1;193(3):757–763. doi: 10.1042/bj1930757. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bisson R., Steffens G. C., Buse G. Localization of lipid binding domain(s) on subunit II of beef heart cytochrome c oxidase. J Biol Chem. 1982 Jun 25;257(12):6716–6720. [PubMed] [Google Scholar]
- Black J. D., Dolly J. O. Interaction of 125I-labeled botulinum neurotoxins with nerve terminals. I. Ultrastructural autoradiographic localization and quantitation of distinct membrane acceptors for types A and B on motor nerves. J Cell Biol. 1986 Aug;103(2):521–534. doi: 10.1083/jcb.103.2.521. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Blaustein R. O., Germann W. J., Finkelstein A., DasGupta B. R. The N-terminal half of the heavy chain of botulinum type A neurotoxin forms channels in planar phospholipid bilayers. FEBS Lett. 1987 Dec 21;226(1):115–120. doi: 10.1016/0014-5793(87)80562-8. [DOI] [PubMed] [Google Scholar]
- Blewitt M. G., Chung L. A., London E. Effect of pH on the conformation of diphtheria toxin and its implications for membrane penetration. Biochemistry. 1985 Sep 24;24(20):5458–5464. doi: 10.1021/bi00341a027. [DOI] [PubMed] [Google Scholar]
- DasGupta B. R., Rasmussen S. Purification and amino acid composition of type E botulinum neurotoxin. Toxicon. 1983;21(4):535–545. doi: 10.1016/0041-0101(83)90131-9. [DOI] [PubMed] [Google Scholar]
- DasGupta B. R., Sathyamoorthy V. Purification and amino acid composition of type A botulinum neurotoxin. Toxicon. 1984;22(3):415–424. doi: 10.1016/0041-0101(84)90085-0. [DOI] [PubMed] [Google Scholar]
- DasGupta B. R., Woody M. A. Amino acid composition of Clostridium botulinum type B neurotoxin. Toxicon. 1984;22(2):312–315. doi: 10.1016/0041-0101(84)90033-3. [DOI] [PubMed] [Google Scholar]
- Dolly J. O., Black J., Williams R. S., Melling J. Acceptors for botulinum neurotoxin reside on motor nerve terminals and mediate its internalization. Nature. 1984 Feb 2;307(5950):457–460. doi: 10.1038/307457a0. [DOI] [PubMed] [Google Scholar]
- Donovan J. J., Middlebrook J. L. Ion-conducting channels produced by botulinum toxin in planar lipid membranes. Biochemistry. 1986 May 20;25(10):2872–2876. doi: 10.1021/bi00358a020. [DOI] [PubMed] [Google Scholar]
- Evans D. M., Williams R. S., Shone C. C., Hambleton P., Melling J., Dolly J. O. Botulinum neurotoxin type B. Its purification, radioiodination and interaction with rat-brain synaptosomal membranes. Eur J Biochem. 1986 Jan 15;154(2):409–416. doi: 10.1111/j.1432-1033.1986.tb09413.x. [DOI] [PubMed] [Google Scholar]
- Giraudat J., Montecucco C., Bisson R., Changeux J. P. Transmembrane topology of acetylcholine receptor subunits probed with photoreactive phospholipids. Biochemistry. 1985 Jun 18;24(13):3121–3127. doi: 10.1021/bi00334a008. [DOI] [PubMed] [Google Scholar]
- Habermann E., Dreyer F. Clostridial neurotoxins: handling and action at the cellular and molecular level. Curr Top Microbiol Immunol. 1986;129:93–179. doi: 10.1007/978-3-642-71399-6_2. [DOI] [PubMed] [Google Scholar]
- Hoch D. H., Romero-Mira M., Ehrlich B. E., Finkelstein A., DasGupta B. R., Simpson L. L. Channels formed by botulinum, tetanus, and diphtheria toxins in planar lipid bilayers: relevance to translocation of proteins across membranes. Proc Natl Acad Sci U S A. 1985 Mar;82(6):1692–1696. doi: 10.1073/pnas.82.6.1692. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hoppe J., Montecucco C., Friedl P. Labeling of subunit b of the ATP synthase from Escherichia coli with a photoreactive phospholipid analogue. J Biol Chem. 1983 Mar 10;258(5):2882–2885. [PubMed] [Google Scholar]
- Jones D. H., Matus A. I. Isolation of synaptic plasma membrane from brain by combined flotation-sedimentation density gradient centrifugation. Biochim Biophys Acta. 1974 Aug 9;356(3):276–287. doi: 10.1016/0005-2736(74)90268-5. [DOI] [PubMed] [Google Scholar]
- Kamata Y., Kozaki S., Sakaguchi G., Iwamori M., Nagai Y. Evidence for direct binding of Clostridium botulinum type E derivative toxin and its fragments to gangliosides and free fatty acids. Biochem Biophys Res Commun. 1986 Nov 14;140(3):1015–1019. doi: 10.1016/0006-291x(86)90736-9. [DOI] [PubMed] [Google Scholar]
- Kitamura M., Iwamori M., Nagai Y. Interaction between Clostridium botulinum neurotoxin and gangliosides. Biochim Biophys Acta. 1980 Mar 20;628(3):328–335. doi: 10.1016/0304-4165(80)90382-7. [DOI] [PubMed] [Google Scholar]
- Markwell M. A., Haas S. M., Tolbert N. E., Bieber L. L. Protein determination in membrane and lipoprotein samples: manual and automated procedures. Methods Enzymol. 1981;72:296–303. doi: 10.1016/s0076-6879(81)72018-4. [DOI] [PubMed] [Google Scholar]
- Mellman I., Fuchs R., Helenius A. Acidification of the endocytic and exocytic pathways. Annu Rev Biochem. 1986;55:663–700. doi: 10.1146/annurev.bi.55.070186.003311. [DOI] [PubMed] [Google Scholar]
- Montecucco C., Bisson R., Dabbeni-Sala F., Pitotti A., Gutweniger H. Interaction of the mitochondrial ATPase complex with phospholipids. J Biol Chem. 1980 Nov 10;255(21):10040–10041. [PubMed] [Google Scholar]
- Montecucco C., Dabbeni-Sala F., Friedl P., Galante Y. M. Membrane topology of ATP synthase from bovine heart mitochondria and Escherichia coli. Eur J Biochem. 1983 Apr 15;132(1):189–194. doi: 10.1111/j.1432-1033.1983.tb07346.x. [DOI] [PubMed] [Google Scholar]
- Montecucco C. Photoreactive lipids for the study of membrane-penetrating toxins. Methods Enzymol. 1988;165:347–357. doi: 10.1016/s0076-6879(88)65052-x. [DOI] [PubMed] [Google Scholar]
- Montecucco C., Schiavo G., Brunner J., Duflot E., Boquet P., Roa M. Tetanus toxin is labeled with photoactivatable phospholipids at low pH. Biochemistry. 1986 Feb 25;25(4):919–924. doi: 10.1021/bi00352a027. [DOI] [PubMed] [Google Scholar]
- Montecucco C., Schiavo G., Gao Z., Bauerlein E., Boquet P., DasGupta B. R. Interaction of botulinum and tetanus toxins with the lipid bilayer surface. Biochem J. 1988 Apr 15;251(2):379–383. doi: 10.1042/bj2510379. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Montecucco C., Schiavo G., Tomasi M. pH-dependence of the phospholipid interaction of diphtheria-toxin fragments. Biochem J. 1985 Oct 1;231(1):123–128. doi: 10.1042/bj2310123. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ochanda J. O., Syuto B., Ohishi I., Naiki M., Kubo S. Binding of Clostridium botulinum neurotoxin to gangliosides. J Biochem. 1986 Jul;100(1):27–33. doi: 10.1093/oxfordjournals.jbchem.a121702. [DOI] [PubMed] [Google Scholar]
- Papini E., Sandoná D., Rappuoli R., Montecucco C. On the membrane translocation of diphtheria toxin: at low pH the toxin induces ion channels on cells. EMBO J. 1988 Nov;7(11):3353–3359. doi: 10.1002/j.1460-2075.1988.tb03207.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Papini E., Schiavo G., Tomasi M., Colombatti M., Rappuoli R., Montecucco C. Lipid interaction of diphtheria toxin and mutants with altered fragment B. 2. Hydrophobic photolabelling and cell intoxication. Eur J Biochem. 1987 Dec 15;169(3):637–644. doi: 10.1111/j.1432-1033.1987.tb13655.x. [DOI] [PubMed] [Google Scholar]
- Ribi H. O., Ludwig D. S., Mercer K. L., Schoolnik G. K., Kornberg R. D. Three-dimensional structure of cholera toxin penetrating a lipid membrane. Science. 1988 Mar 11;239(4845):1272–1276. doi: 10.1126/science.3344432. [DOI] [PubMed] [Google Scholar]
- Sakaguchi G. Clostridium botulinum toxins. Pharmacol Ther. 1982;19(2):165–194. doi: 10.1016/0163-7258(82)90061-4. [DOI] [PubMed] [Google Scholar]
- Sathyamoorthy V., DasGupta B. R. Separation, purification, partial characterization and comparison of the heavy and light chains of botulinum neurotoxin types A, B, and E. J Biol Chem. 1985 Sep 5;260(19):10461–10466. [PubMed] [Google Scholar]
- Shone C. C., Hambleton P., Melling J. A 50-kDa fragment from the NH2-terminus of the heavy subunit of Clostridium botulinum type A neurotoxin forms channels in lipid vesicles. Eur J Biochem. 1987 Aug 17;167(1):175–180. doi: 10.1111/j.1432-1033.1987.tb13320.x. [DOI] [PubMed] [Google Scholar]
- Simpson L. L. Ammonium chloride and methylamine hydrochloride antagonize clostridial neurotoxins. J Pharmacol Exp Ther. 1983 Jun;225(3):546–552. [PubMed] [Google Scholar]
- Simpson L. L. Molecular pharmacology of botulinum toxin and tetanus toxin. Annu Rev Pharmacol Toxicol. 1986;26:427–453. doi: 10.1146/annurev.pa.26.040186.002235. [DOI] [PubMed] [Google Scholar]
- Simpson L. L., Rapport M. M. The binding of botulinum toxin to membrane lipids: sphingolipids, steroids and fatty acids. J Neurochem. 1971 Sep;18(9):1751–1759. doi: 10.1111/j.1471-4159.1971.tb03750.x. [DOI] [PubMed] [Google Scholar]
- Simpson L. L. The interaction between aminoquinolines and presynaptically acting neurotoxins. J Pharmacol Exp Ther. 1982 Jul;222(1):43–48. [PubMed] [Google Scholar]
- Sugiyama H. Clostridium botulinum neurotoxin. Microbiol Rev. 1980 Sep;44(3):419–448. doi: 10.1128/mr.44.3.419-448.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tomasi M., Montecucco C. Lipid insertion of cholera toxin after binding to GM1-containing liposomes. J Biol Chem. 1981 Nov 10;256(21):11177–11181. [PubMed] [Google Scholar]
- Williams R. S., Tse C. K., Dolly J. O., Hambleton P., Melling J. Radioiodination of botulinum neurotoxin type A with retention of biological activity and its binding to brain synaptosomes. Eur J Biochem. 1983 Mar 15;131(2):437–445. doi: 10.1111/j.1432-1033.1983.tb07282.x. [DOI] [PubMed] [Google Scholar]