Home LiteratureArticle Details
PMID: 8520474 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Formation of ion channels in lipid bilayers by a peptide with the predicted transmembrane sequence of botulinum neurotoxin A.

Protein science : a publication of the Protein Society ·Vol. 4 ·No. 8 ·1995-08-00 ·Pages 1490-7

Oblatt-Montal M, Yamazaki M, Nelson R, Montal M

Abstract

Synthetic peptides patterned after the predicted transmembrane sequence of botulinum toxin A were used as tools to identify an ion channel-forming motif. A peptide denoted BoTxATM, with the sequence GAVILLEFIPEIAI PVLGTFALV, forms cation-selective channels when reconstituted in planar lipid bilayers. As predicted, the self-assembled conductive oligomers express heterogeneous single-channel conductances. The most frequent openings exhibit single-channel conductance of 12 and 7 pS in 0.5 M NaCl, and 29 and 9 pS in 0.5 M KCl. In contrast, ion channels are not formed by a peptide of the same amino acid composition as BoTxATM with a scrambled sequence. Conformational energy calculations show that a bundle of four amphipathic alpha-helices is a plausible structural motif underlying the measured pore properties. These studies suggest that the identified module may play a functional role in the ion channel-forming activity of intact botulinum toxin A.

MeSH Terms
Amino Acid Sequence Botulinum Toxins/chemistry Cell Membrane/chemistry Hydrogen-Ion Concentration Ion Channels Lipid Bilayers Molecular Sequence Data Sequence Homology, Amino Acid Tetanus Toxin/chemistry
Chemicals
Ion Channels Lipid Bilayers Tetanus Toxin Botulinum Toxins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Oblatt-Montal M
Department of Biology, University of California San Diego, La Jolla 92093-0366, USA.
Yamazaki M
Nelson R
Montal M
References (46)
46 references, click to expand
  1. Tetanus toxin fragment forms channels in lipid vesicles at low pH.
    Proc Natl Acad Sci U S A. 1982 Dec;79(24):7614-8 PMID: 6296842
  2. Synaptic vesicles and exocytosis.
    Annu Rev Neurosci. 1994;17:219-46 PMID: 8210174
  3. Synthetic peptides and four-helix bundle proteins as model systems for the pore-forming structure of channel proteins. I. Transmembrane segment M2 of the nicotinic cholinergic receptor channel is a key pore-lining structure.
    J Biol Chem. 1993 Jul 15;268(20):14601-7 PMID: 7686900
  4. Identification of the nerve terminal targets of botulinum neurotoxin serotypes A, D, and E.
    J Biol Chem. 1993 Nov 15;268(32):23784-7 PMID: 8226912
  5. Botulinum neurotoxins are zinc proteins.
    J Biol Chem. 1992 Nov 25;267(33):23479-83 PMID: 1429690
  6. Crystal structure of cholera toxin B-pentamer bound to receptor GM1 pentasaccharide.
    Protein Sci. 1994 Feb;3(2):166-75 PMID: 8003954
  7. Synthetic peptides and proteins as models for pore-forming structure of channel proteins.
    Methods Enzymol. 1992;207:510-25 PMID: 1382200
  8. Acetylcholine receptor channel imaged in the open state.
    Nature. 1995 Jan 5;373(6509):37-43 PMID: 7800037
  9. Predicted secondary structure of the 20 S proteasome and model structure of the putative peptide channel.
    FEBS Lett. 1994 Oct 31;354(1):45-9 PMID: 7957899
  10. A protein-conducting channel in the endoplasmic reticulum.
    Cell. 1991 May 3;65(3):371-80 PMID: 1902142
  11. M2 delta, a candidate for the structure lining the ionic channel of the nicotinic cholinergic receptor.
    Proc Natl Acad Sci U S A. 1988 Nov;85(22):8703-7 PMID: 2460876
  12. Synaptic vesicle membrane fusion complex: action of clostridial neurotoxins on assembly.
    EMBO J. 1994 Nov 1;13(21):5051-61 PMID: 7957071
  13. Signal peptides open protein-conducting channels in E. coli.
    Cell. 1992 May 15;69(4):677-84 PMID: 1375130
  14. Interaction of Clostridium botulinum C2 toxin with lipid bilayer membranes. Formation of cation-selective channels and inhibition of channel function by chloroquine.
    J Biol Chem. 1994 Jun 17;269(24):16706-11 PMID: 7515883
  15. High-resolution structures of photosynthetic reaction centers.
    Annu Rev Biophys Biophys Chem. 1991;20:247-66 PMID: 1867718
  16. Progressive alignment and phylogenetic tree construction of protein sequences.
    Methods Enzymol. 1990;183:375-87 PMID: 2314283
  17. A mechanism for ion selectivity in potassium channels: computational studies of cation-pi interactions.
    Science. 1993 Sep 24;261(5129):1708-10 PMID: 8378771
  18. Botulinum neurotoxin C1 blocks neurotransmitter release by means of cleaving HPC-1/syntaxin.
    EMBO J. 1993 Dec;12(12):4821-8 PMID: 7901002
  19. Tetanus toxin forms channels in planar lipid bilayers containing gangliosides.
    Biophys J. 1984 Jan;45(1):83-5 PMID: 19431576
  20. Mechanism of action of tetanus and botulinum neurotoxins.
    Mol Microbiol. 1994 Jul;13(1):1-8 PMID: 7527117
  21. A molecular description of synaptic vesicle membrane trafficking.
    Annu Rev Biochem. 1994;63:63-100 PMID: 7979250
  22. Analysis of membrane and surface protein sequences with the hydrophobic moment plot.
    J Mol Biol. 1984 Oct 15;179(1):125-42 PMID: 6502707
  23. The pore dimensions of gramicidin A.
    Biophys J. 1993 Dec;65(6):2455-60 PMID: 7508762
  24. Formation of bimolecular membranes from lipid monolayers and a study of their electrical properties.
    Proc Natl Acad Sci U S A. 1972 Dec;69(12):3561-6 PMID: 4509315
  25. Ion-conducting channels produced by botulinum toxin in planar lipid membranes.
    Biochemistry. 1986 May 20;25(10):2872-6 PMID: 2424493
  26. Botulinum neurotoxins serotypes A and E cleave SNAP-25 at distinct COOH-terminal peptide bonds.
    FEBS Lett. 1993 Nov 29;335(1):99-103 PMID: 8243676
  27. 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):115-20 PMID: 2446925
  28. Kinetic analysis of channel gating. Application to the cholinergic receptor channel and the chloride channel from Torpedo californica.
    Biophys J. 1985 Apr;47(4):469-78 PMID: 2580569
  29. Mitochondrial protein import: reversible binding of the presequence at the trans side of the outer membrane drives partial translocation and unfolding.
    Cell. 1995 Jan 13;80(1):127-37 PMID: 7813008
  30. Nicotinic acetylcholine receptor at 9 A resolution.
    J Mol Biol. 1993 Feb 20;229(4):1101-24 PMID: 8445638
  31. Single-channel recordings from purified acetylcholine receptors reconstituted in bilayers formed at the tip of patch pipets.
    Biochemistry. 1983 May 10;22(10):2319-23 PMID: 6305400
  32. Refined structure of Escherichia coli heat-labile enterotoxin, a close relative of cholera toxin.
    J Mol Biol. 1993 Apr 5;230(3):890-918 PMID: 8478941
  33. Identification of an ion channel-forming motif in the primary structure of tetanus and botulinum neurotoxins.
    FEBS Lett. 1992 Nov 16;313(1):12-8 PMID: 1385218
  34. Characterization of the channel properties of tetanus toxin in planar lipid bilayers.
    Biophys J. 1988 May;53(5):771-83 PMID: 2455552
  35. Bundles of amphipathic transmembrane alpha-helices as a structural motif for ion-conducting channel proteins: studies on sodium channels and acetylcholine receptors.
    Proteins. 1990;8(3):226-36 PMID: 2177892
  36. Crystal structure of the cell-binding B oligomer of verotoxin-1 from E. coli.
    Nature. 1992 Feb 20;355(6362):748-50 PMID: 1741063
  37. Bacterial protein toxins penetrate cells via a four-step mechanism.
    FEBS Lett. 1994 Jun 6;346(1):92-8 PMID: 8206166
  38. Pore formation by tetanus toxin, its chain and fragments in neuronal membranes and evaluation of the underlying motifs in the structure of the toxin molecule.
    Naunyn Schmiedebergs Arch Pharmacol. 1994 Jan;349(1):66-73 PMID: 8139702
  39. Low pH induces a hydrophobic domain in the tetanus toxin molecule.
    Eur J Biochem. 1984 Oct 15;144(2):339-44 PMID: 6489333
  40. 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):1692-6 PMID: 3856850
  41. Synporins--synthetic proteins that emulate the pore structure of biological ionic channels.
    Proc Natl Acad Sci U S A. 1990 Sep;87(18):6929-33 PMID: 1698285
  42. Channels formed in phospholipid bilayer membranes by diphtheria, tetanus, botulinum and anthrax toxin.
    J Physiol (Paris). 1990;84(2):188-90 PMID: 1705290
  43. Tetanus and botulinum-B neurotoxins block neurotransmitter release by proteolytic cleavage of synaptobrevin.
    Nature. 1992 Oct 29;359(6398):832-5 PMID: 1331807
  44. In situ neutralization in Boc-chemistry solid phase peptide synthesis. Rapid, high yield assembly of difficult sequences.
    Int J Pept Protein Res. 1992 Sep-Oct;40(3-4):180-93 PMID: 1478777
  45. Cleavage of members of the synaptobrevin/VAMP family by types D and F botulinal neurotoxins and tetanus toxin.
    J Biol Chem. 1994 Apr 29;269(17):12764-72 PMID: 8175689
  46. The structure of the voltage-sensitive sodium channel. Inferences derived from computer-aided analysis of the Electrophorus electricus channel primary structure.
    FEBS Lett. 1985 Dec 2;193(2):125-34 PMID: 2415395
Article Info
Journal
Protein science : a publication of the Protein Society
Abbr.
Protein Sci
ISSN
0961-8368
DOI
10.1002/pro.5560040806
Published
1995-08-00
Pages
1490-7
Language
English
Region
United States
NLM ID
9211750
PMCID
PMC2143195
Subset
IM
Grants
NIGMS NIH HHS · NIH-5T32GM08326 · United States
External Links
PubMed source DOI: 10.1002/pro.5560040806 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2