Identification of the receptor-binding sites in the carboxyl-terminal half of the heavy chain of botulinum neurotoxin types C and D

Kentaro Tsukamoto, Yuiko Kozai, Hideshi Ihara, Tomoko Kohda, Masafumi Mukamoto, Takao Tsuji, Shunji Kozaki

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Botulinum neurotoxin (BoNT) binds to presynaptic neuronal cells and blocks neurotransmitter release. The carboxyl-terminal half of the heavy chain (HC) of the neurotoxin recognizes its specific receptor on the plasma membrane. We have previously demonstrated that BoNT/C binds to gangliosides GD1b and GT1b under physiological conditions, while BoNT/D interacts with phosphatidylethanolamine (PE). Here we report that the recognition sites for gangliosides and PE are present in the carboxyl-terminal domain of HC. Chimeric mutants and site-directed mutants of BoNT/C-HC and BoNT/D-HC were generated and their binding activities evaluated. The chimeric HC that consisted of the amino-terminal half of BoNT/D-HC and the carboxyl-terminal half of BoNT/C-HC possessed activity similar to the authentic BoNT/C-HC, suggesting that the carboxyl-terminal region of HC is involved in the receptor recognition of BoNT/C. Moreover, analysis using site-directed mutants indicated that the peptide motif W1257Y⋯G1270⋯H1282 plays an important role in the interaction between BoNT/C and gangliosides. In contrast, we revealed that two lysine residues of BoNT/D-HC are involved in the formation of the critical binding site for receptor binding.

Original languageEnglish
Pages (from-to)484-493
Number of pages10
JournalMicrobial Pathogenesis
Volume44
Issue number6
DOIs
Publication statusPublished - 01-06-2008

Fingerprint

Binding Sites
Gangliosides
Neurotoxins
Lysine
Neurotransmitter Agents
botulinum toxin type D
botulinum toxin type C
Cell Membrane
Peptides
phosphatidylethanolamine

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Infectious Diseases

Cite this

Tsukamoto, Kentaro ; Kozai, Yuiko ; Ihara, Hideshi ; Kohda, Tomoko ; Mukamoto, Masafumi ; Tsuji, Takao ; Kozaki, Shunji. / Identification of the receptor-binding sites in the carboxyl-terminal half of the heavy chain of botulinum neurotoxin types C and D. In: Microbial Pathogenesis. 2008 ; Vol. 44, No. 6. pp. 484-493.
@article{9b2a12084b644c3fa058fd7eaa4cd379,
title = "Identification of the receptor-binding sites in the carboxyl-terminal half of the heavy chain of botulinum neurotoxin types C and D",
abstract = "Botulinum neurotoxin (BoNT) binds to presynaptic neuronal cells and blocks neurotransmitter release. The carboxyl-terminal half of the heavy chain (HC) of the neurotoxin recognizes its specific receptor on the plasma membrane. We have previously demonstrated that BoNT/C binds to gangliosides GD1b and GT1b under physiological conditions, while BoNT/D interacts with phosphatidylethanolamine (PE). Here we report that the recognition sites for gangliosides and PE are present in the carboxyl-terminal domain of HC. Chimeric mutants and site-directed mutants of BoNT/C-HC and BoNT/D-HC were generated and their binding activities evaluated. The chimeric HC that consisted of the amino-terminal half of BoNT/D-HC and the carboxyl-terminal half of BoNT/C-HC possessed activity similar to the authentic BoNT/C-HC, suggesting that the carboxyl-terminal region of HC is involved in the receptor recognition of BoNT/C. Moreover, analysis using site-directed mutants indicated that the peptide motif W1257Y⋯G1270⋯H1282 plays an important role in the interaction between BoNT/C and gangliosides. In contrast, we revealed that two lysine residues of BoNT/D-HC are involved in the formation of the critical binding site for receptor binding.",
author = "Kentaro Tsukamoto and Yuiko Kozai and Hideshi Ihara and Tomoko Kohda and Masafumi Mukamoto and Takao Tsuji and Shunji Kozaki",
year = "2008",
month = "6",
day = "1",
doi = "10.1016/j.micpath.2007.12.003",
language = "English",
volume = "44",
pages = "484--493",
journal = "Microbial Pathogenesis",
issn = "0882-4010",
publisher = "Academic Press Inc.",
number = "6",

}

Identification of the receptor-binding sites in the carboxyl-terminal half of the heavy chain of botulinum neurotoxin types C and D. / Tsukamoto, Kentaro; Kozai, Yuiko; Ihara, Hideshi; Kohda, Tomoko; Mukamoto, Masafumi; Tsuji, Takao; Kozaki, Shunji.

In: Microbial Pathogenesis, Vol. 44, No. 6, 01.06.2008, p. 484-493.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Identification of the receptor-binding sites in the carboxyl-terminal half of the heavy chain of botulinum neurotoxin types C and D

AU - Tsukamoto, Kentaro

AU - Kozai, Yuiko

AU - Ihara, Hideshi

AU - Kohda, Tomoko

AU - Mukamoto, Masafumi

AU - Tsuji, Takao

AU - Kozaki, Shunji

PY - 2008/6/1

Y1 - 2008/6/1

N2 - Botulinum neurotoxin (BoNT) binds to presynaptic neuronal cells and blocks neurotransmitter release. The carboxyl-terminal half of the heavy chain (HC) of the neurotoxin recognizes its specific receptor on the plasma membrane. We have previously demonstrated that BoNT/C binds to gangliosides GD1b and GT1b under physiological conditions, while BoNT/D interacts with phosphatidylethanolamine (PE). Here we report that the recognition sites for gangliosides and PE are present in the carboxyl-terminal domain of HC. Chimeric mutants and site-directed mutants of BoNT/C-HC and BoNT/D-HC were generated and their binding activities evaluated. The chimeric HC that consisted of the amino-terminal half of BoNT/D-HC and the carboxyl-terminal half of BoNT/C-HC possessed activity similar to the authentic BoNT/C-HC, suggesting that the carboxyl-terminal region of HC is involved in the receptor recognition of BoNT/C. Moreover, analysis using site-directed mutants indicated that the peptide motif W1257Y⋯G1270⋯H1282 plays an important role in the interaction between BoNT/C and gangliosides. In contrast, we revealed that two lysine residues of BoNT/D-HC are involved in the formation of the critical binding site for receptor binding.

AB - Botulinum neurotoxin (BoNT) binds to presynaptic neuronal cells and blocks neurotransmitter release. The carboxyl-terminal half of the heavy chain (HC) of the neurotoxin recognizes its specific receptor on the plasma membrane. We have previously demonstrated that BoNT/C binds to gangliosides GD1b and GT1b under physiological conditions, while BoNT/D interacts with phosphatidylethanolamine (PE). Here we report that the recognition sites for gangliosides and PE are present in the carboxyl-terminal domain of HC. Chimeric mutants and site-directed mutants of BoNT/C-HC and BoNT/D-HC were generated and their binding activities evaluated. The chimeric HC that consisted of the amino-terminal half of BoNT/D-HC and the carboxyl-terminal half of BoNT/C-HC possessed activity similar to the authentic BoNT/C-HC, suggesting that the carboxyl-terminal region of HC is involved in the receptor recognition of BoNT/C. Moreover, analysis using site-directed mutants indicated that the peptide motif W1257Y⋯G1270⋯H1282 plays an important role in the interaction between BoNT/C and gangliosides. In contrast, we revealed that two lysine residues of BoNT/D-HC are involved in the formation of the critical binding site for receptor binding.

UR - http://www.scopus.com/inward/record.url?scp=43049163811&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=43049163811&partnerID=8YFLogxK

U2 - 10.1016/j.micpath.2007.12.003

DO - 10.1016/j.micpath.2007.12.003

M3 - Article

VL - 44

SP - 484

EP - 493

JO - Microbial Pathogenesis

JF - Microbial Pathogenesis

SN - 0882-4010

IS - 6

ER -