Specific inhibition of cytotoxicity of Shiga-like toxin 1 of enterohemorrhagic Escherichia coli by gallocatechin gallate and epigallocatechin gallate

Takahisa Miyamoto, Seiyo Toyofuku, Narumi Tachiki, Etsuko Kimura, Ting Zhou, Tadahiro Ozawa, Motokazu Nakayama, Naofumi Shigemune, Kanami Shimatani, Hajime Tokuda, Ken-ichi Honjoh

Research output: Contribution to journalArticle

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Abstract

Mechanism of inhibitory action of 8 catechins and teaflavin was investigated at low concentration against Shiga-like toxin (Stx). Viability of Vero cells largely decreased in the presence of Stx1 and Stx2 preparations. Cytotoxicity of Stx1 decreased after preincubation with gallocatechin gallate (GCg) and epigallocatechin gallate (EGCg) at 100mg/L. However, the cytotoxicity of Stx2 was not inhibited by the preincubation with catechins and teaflavin tested. The inhibitory activity of GCg and EGCg at 15mg/L (0.0327mM) was investigated against Stx preparations with various concentrations. The cytotoxicity of Stx1 at the concentration ranging from 1.6 to 50ng/mL significantly reduced (p<0.01) by the preincubation of Stx1 with GCg at 15mg/L. Similarly, the cytotoxicity of Stx1 at the concentration ranging from 3.1 to 25ng/mL was significantly reduced (p<0.01) by the preincubation with EGCg at 15mg/L. In contrast, GCg and EGCg did not inhibit cytotoxicity of Stx2 at any concentrations tested. On the other hand, EGC showed no significant effects on cytotoxicity of both Stx1 and Stx2 at the same concentrations tested. The pocket sizes formed at the center of the Stx1B and Stx2B pentamers were calculated to be 778Å3 and 475Å3, respectively. Docking simulations were conducted with EGCg positioned in the center of the pore of StxB pentamers. The docking models showed that EGCg formed 7 hydrogen bonds with side chains of amino acids faced inside the pocket of the Stx1B pentarmer with the lowest intramolecular energy (strain energy+electrostatic energy) of-0.1kcal/mol. In contrast, in the case of Stx2B pentamer, EGCg formed 6 hydrogen bonds with the lowest intramolecular energy of 5.2kcal/mol. In silico study suggested that EGCg forms more stable structure with Stx1B pentamer than Stx2B pentamer. These results indicated that both GCg and EGCg specifically inhibited cytotoxicity of Stx1 but not of Stx2.

Original languageEnglish
Pages (from-to)263-269
Number of pages7
JournalFood Control
Volume42
DOIs
Publication statusPublished - Aug 1 2014

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Shiga Toxin 1
Shiga-like toxin 1
Shiga Toxins
Enterohemorrhagic Escherichia coli
enterohemorrhagic Escherichia coli
epigallocatechin
cytotoxicity
Catechin
Hydrogen
Shiga-like toxins
energy
Vero Cells
flavanols
epigallocatechin gallate
gallocatechin gallate
hydrogen
Static Electricity
Computer Simulation
Amino Acids

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Food Science

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Specific inhibition of cytotoxicity of Shiga-like toxin 1 of enterohemorrhagic Escherichia coli by gallocatechin gallate and epigallocatechin gallate. / Miyamoto, Takahisa; Toyofuku, Seiyo; Tachiki, Narumi; Kimura, Etsuko; Zhou, Ting; Ozawa, Tadahiro; Nakayama, Motokazu; Shigemune, Naofumi; Shimatani, Kanami; Tokuda, Hajime; Honjoh, Ken-ichi.

In: Food Control, Vol. 42, 01.08.2014, p. 263-269.

Research output: Contribution to journalArticle

Miyamoto, Takahisa ; Toyofuku, Seiyo ; Tachiki, Narumi ; Kimura, Etsuko ; Zhou, Ting ; Ozawa, Tadahiro ; Nakayama, Motokazu ; Shigemune, Naofumi ; Shimatani, Kanami ; Tokuda, Hajime ; Honjoh, Ken-ichi. / Specific inhibition of cytotoxicity of Shiga-like toxin 1 of enterohemorrhagic Escherichia coli by gallocatechin gallate and epigallocatechin gallate. In: Food Control. 2014 ; Vol. 42. pp. 263-269.
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abstract = "Mechanism of inhibitory action of 8 catechins and teaflavin was investigated at low concentration against Shiga-like toxin (Stx). Viability of Vero cells largely decreased in the presence of Stx1 and Stx2 preparations. Cytotoxicity of Stx1 decreased after preincubation with gallocatechin gallate (GCg) and epigallocatechin gallate (EGCg) at 100mg/L. However, the cytotoxicity of Stx2 was not inhibited by the preincubation with catechins and teaflavin tested. The inhibitory activity of GCg and EGCg at 15mg/L (0.0327mM) was investigated against Stx preparations with various concentrations. The cytotoxicity of Stx1 at the concentration ranging from 1.6 to 50ng/mL significantly reduced (p<0.01) by the preincubation of Stx1 with GCg at 15mg/L. Similarly, the cytotoxicity of Stx1 at the concentration ranging from 3.1 to 25ng/mL was significantly reduced (p<0.01) by the preincubation with EGCg at 15mg/L. In contrast, GCg and EGCg did not inhibit cytotoxicity of Stx2 at any concentrations tested. On the other hand, EGC showed no significant effects on cytotoxicity of both Stx1 and Stx2 at the same concentrations tested. The pocket sizes formed at the center of the Stx1B and Stx2B pentamers were calculated to be 778{\AA}3 and 475{\AA}3, respectively. Docking simulations were conducted with EGCg positioned in the center of the pore of StxB pentamers. The docking models showed that EGCg formed 7 hydrogen bonds with side chains of amino acids faced inside the pocket of the Stx1B pentarmer with the lowest intramolecular energy (strain energy+electrostatic energy) of-0.1kcal/mol. In contrast, in the case of Stx2B pentamer, EGCg formed 6 hydrogen bonds with the lowest intramolecular energy of 5.2kcal/mol. In silico study suggested that EGCg forms more stable structure with Stx1B pentamer than Stx2B pentamer. These results indicated that both GCg and EGCg specifically inhibited cytotoxicity of Stx1 but not of Stx2.",
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AU - Miyamoto, Takahisa

AU - Toyofuku, Seiyo

AU - Tachiki, Narumi

AU - Kimura, Etsuko

AU - Zhou, Ting

AU - Ozawa, Tadahiro

AU - Nakayama, Motokazu

AU - Shigemune, Naofumi

AU - Shimatani, Kanami

AU - Tokuda, Hajime

AU - Honjoh, Ken-ichi

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N2 - Mechanism of inhibitory action of 8 catechins and teaflavin was investigated at low concentration against Shiga-like toxin (Stx). Viability of Vero cells largely decreased in the presence of Stx1 and Stx2 preparations. Cytotoxicity of Stx1 decreased after preincubation with gallocatechin gallate (GCg) and epigallocatechin gallate (EGCg) at 100mg/L. However, the cytotoxicity of Stx2 was not inhibited by the preincubation with catechins and teaflavin tested. The inhibitory activity of GCg and EGCg at 15mg/L (0.0327mM) was investigated against Stx preparations with various concentrations. The cytotoxicity of Stx1 at the concentration ranging from 1.6 to 50ng/mL significantly reduced (p<0.01) by the preincubation of Stx1 with GCg at 15mg/L. Similarly, the cytotoxicity of Stx1 at the concentration ranging from 3.1 to 25ng/mL was significantly reduced (p<0.01) by the preincubation with EGCg at 15mg/L. In contrast, GCg and EGCg did not inhibit cytotoxicity of Stx2 at any concentrations tested. On the other hand, EGC showed no significant effects on cytotoxicity of both Stx1 and Stx2 at the same concentrations tested. The pocket sizes formed at the center of the Stx1B and Stx2B pentamers were calculated to be 778Å3 and 475Å3, respectively. Docking simulations were conducted with EGCg positioned in the center of the pore of StxB pentamers. The docking models showed that EGCg formed 7 hydrogen bonds with side chains of amino acids faced inside the pocket of the Stx1B pentarmer with the lowest intramolecular energy (strain energy+electrostatic energy) of-0.1kcal/mol. In contrast, in the case of Stx2B pentamer, EGCg formed 6 hydrogen bonds with the lowest intramolecular energy of 5.2kcal/mol. In silico study suggested that EGCg forms more stable structure with Stx1B pentamer than Stx2B pentamer. These results indicated that both GCg and EGCg specifically inhibited cytotoxicity of Stx1 but not of Stx2.

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