The N-terminal Arg residue is essential for autocatalytic activation of a lipopolysaccharide-responsive protease zymogen

Yuki Kobayashi, Takafumi Shiga, Toshio Shibata, Miyuki Sako, Katsumi Maenaka, Takumi Koshiba, Hikaru Mizumura, Toshio Oda, Shun-Ichiro Kawabata

研究成果: ジャーナルへの寄稿記事

8 引用 (Scopus)

抄録

Factor C, a serine protease zymogen involved in innate immune responses in horseshoe crabs, is known to be autocatalytically activated on the surface of bacterial lipopolysaccharides, but the molecular mechanism of this activation remains unknown. In this study, we show that wild-type factor C expressed in HEK293S cells exhibits a lipopolysaccharide-induced activity equivalent to that of native factor C. Analysis of the N-terminal addition, deletion, or substitution mutants shows that the N-terminal Arg residue and the distance between the N terminus and the tripartite of lipopolysaccharide-binding site are essential factors for autocatalytic activation, and that the positive charge of theNterminus may interact with an acidic amino acid(s) of the molecule to convert the zymogen into an active form. Chemical cross-linking experiments indicate that the N terminus is required to form a complex of the factor C molecules in a sufficiently close vicinity to be chemically cross-linked on the surface of lipopolysaccharides. We propose a molecular mechanism of the autocatalytic activation of the protease zymogen on lipopolysaccharides functioning as a platform to induce specific protein-protein interaction between the factor C molecules.

元の言語英語
ページ(範囲)25987-25995
ページ数9
ジャーナルJournal of Biological Chemistry
289
発行部数37
DOI
出版物ステータス出版済み - 9 12 2014

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Enzyme Precursors
Lipopolysaccharides
Peptide Hydrolases
Chemical activation
Molecules
Acidic Amino Acids
Horseshoe Crabs
Serine Proteases
Innate Immunity
Statistical Factor Analysis
Proteins
Substitution reactions
Binding Sites
Experiments

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

これを引用

The N-terminal Arg residue is essential for autocatalytic activation of a lipopolysaccharide-responsive protease zymogen. / Kobayashi, Yuki; Shiga, Takafumi; Shibata, Toshio; Sako, Miyuki; Maenaka, Katsumi; Koshiba, Takumi; Mizumura, Hikaru; Oda, Toshio; Kawabata, Shun-Ichiro.

:: Journal of Biological Chemistry, 巻 289, 番号 37, 12.09.2014, p. 25987-25995.

研究成果: ジャーナルへの寄稿記事

Kobayashi, Yuki ; Shiga, Takafumi ; Shibata, Toshio ; Sako, Miyuki ; Maenaka, Katsumi ; Koshiba, Takumi ; Mizumura, Hikaru ; Oda, Toshio ; Kawabata, Shun-Ichiro. / The N-terminal Arg residue is essential for autocatalytic activation of a lipopolysaccharide-responsive protease zymogen. :: Journal of Biological Chemistry. 2014 ; 巻 289, 番号 37. pp. 25987-25995.
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abstract = "Factor C, a serine protease zymogen involved in innate immune responses in horseshoe crabs, is known to be autocatalytically activated on the surface of bacterial lipopolysaccharides, but the molecular mechanism of this activation remains unknown. In this study, we show that wild-type factor C expressed in HEK293S cells exhibits a lipopolysaccharide-induced activity equivalent to that of native factor C. Analysis of the N-terminal addition, deletion, or substitution mutants shows that the N-terminal Arg residue and the distance between the N terminus and the tripartite of lipopolysaccharide-binding site are essential factors for autocatalytic activation, and that the positive charge of theNterminus may interact with an acidic amino acid(s) of the molecule to convert the zymogen into an active form. Chemical cross-linking experiments indicate that the N terminus is required to form a complex of the factor C molecules in a sufficiently close vicinity to be chemically cross-linked on the surface of lipopolysaccharides. We propose a molecular mechanism of the autocatalytic activation of the protease zymogen on lipopolysaccharides functioning as a platform to induce specific protein-protein interaction between the factor C molecules.",
author = "Yuki Kobayashi and Takafumi Shiga and Toshio Shibata and Miyuki Sako and Katsumi Maenaka and Takumi Koshiba and Hikaru Mizumura and Toshio Oda and Shun-Ichiro Kawabata",
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AU - Kobayashi, Yuki

AU - Shiga, Takafumi

AU - Shibata, Toshio

AU - Sako, Miyuki

AU - Maenaka, Katsumi

AU - Koshiba, Takumi

AU - Mizumura, Hikaru

AU - Oda, Toshio

AU - Kawabata, Shun-Ichiro

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N2 - Factor C, a serine protease zymogen involved in innate immune responses in horseshoe crabs, is known to be autocatalytically activated on the surface of bacterial lipopolysaccharides, but the molecular mechanism of this activation remains unknown. In this study, we show that wild-type factor C expressed in HEK293S cells exhibits a lipopolysaccharide-induced activity equivalent to that of native factor C. Analysis of the N-terminal addition, deletion, or substitution mutants shows that the N-terminal Arg residue and the distance between the N terminus and the tripartite of lipopolysaccharide-binding site are essential factors for autocatalytic activation, and that the positive charge of theNterminus may interact with an acidic amino acid(s) of the molecule to convert the zymogen into an active form. Chemical cross-linking experiments indicate that the N terminus is required to form a complex of the factor C molecules in a sufficiently close vicinity to be chemically cross-linked on the surface of lipopolysaccharides. We propose a molecular mechanism of the autocatalytic activation of the protease zymogen on lipopolysaccharides functioning as a platform to induce specific protein-protein interaction between the factor C molecules.

AB - Factor C, a serine protease zymogen involved in innate immune responses in horseshoe crabs, is known to be autocatalytically activated on the surface of bacterial lipopolysaccharides, but the molecular mechanism of this activation remains unknown. In this study, we show that wild-type factor C expressed in HEK293S cells exhibits a lipopolysaccharide-induced activity equivalent to that of native factor C. Analysis of the N-terminal addition, deletion, or substitution mutants shows that the N-terminal Arg residue and the distance between the N terminus and the tripartite of lipopolysaccharide-binding site are essential factors for autocatalytic activation, and that the positive charge of theNterminus may interact with an acidic amino acid(s) of the molecule to convert the zymogen into an active form. Chemical cross-linking experiments indicate that the N terminus is required to form a complex of the factor C molecules in a sufficiently close vicinity to be chemically cross-linked on the surface of lipopolysaccharides. We propose a molecular mechanism of the autocatalytic activation of the protease zymogen on lipopolysaccharides functioning as a platform to induce specific protein-protein interaction between the factor C molecules.

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