Characterization and Properties of a 1,3-β-D-Glucan Pattern Recognition Protein of Tenebrio molitor Larvae That Is Specifically Degraded by Serine Protease during Prophenoloxidase Activation

Rong Zhang, Hae Yun Cho, Hyun Sic Kim, Young Gerl Ma, Tsukasa Osaki, Shun-Ichiro Kawabata, Kenneth Söderhäll, Bok Luel Lee

研究成果: ジャーナルへの寄稿記事

66 引用 (Scopus)

抄録

Although many different pattern recognition receptors recognizing peptidoglycan and 1,3-β-D-glucan have been identified in vertebrates and insects, the molecular mechanism of these molecules in the pattern recognition and subsequent signaling is largely unknown. To gain insights into the action mechanism of 1,3-β-D-glucan pattern recognition protein in the insect prophenoloxidase (proPO) activation system, we purified a 53-kDa 1,3-β-D-glucan recognition protein (Tm-GRP) to homogeneity from the hemolymph of the mealworm, Tenebrio molitor, by using a 1,3-β-D-glucan affinity column. The purified protein specifically bound to 1,3-β-D-glucan but not to peptidoglycan. Subsequent molecular cloning revealed that Tm-GRP contains a region with close sequence similarity to bacterial glucanases. Strikingly, two catalytically important residues in glucanases are replaced with other nonhomologous amino acids in Tm-GRP. The finding suggests that Tm-GRP has evolved from an ancestral gene of glucanases but retained only the ability to recognize 1,3-β-D-glucan. A Western blot analysis of the protein level of endogenous Tm-GRP showed that the protein was specifically degraded following the activation of proPO with 1,3-β-D-glucan and calcium ion. The degradation was significantly retarded by the addition of serine protease inhibitors but not by cysteine or acidic protease inhibitor. These results suggest that 1,3-β-D-glucan pattern recognition protein is specifically degraded by serine protease(s) during proPO activation, and we propose that this degradation is an important regulatory mechanism of the activation of the proPO system.

元の言語英語
ページ(範囲)42072-42079
ページ数8
ジャーナルJournal of Biological Chemistry
278
発行部数43
DOI
出版物ステータス出版済み - 10 24 2003

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Tenebrio
Serine Proteases
Pattern recognition
Larva
Chemical activation
Proteins
Peptidoglycan
Insect Proteins
Pattern Recognition Receptors
Degradation
Serine Proteinase Inhibitors
Hemolymph
pro-phenoloxidase
polyglucosan
Cloning
Molecular Cloning
Protease Inhibitors
Cysteine
Insects
Vertebrates

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

これを引用

Characterization and Properties of a 1,3-β-D-Glucan Pattern Recognition Protein of Tenebrio molitor Larvae That Is Specifically Degraded by Serine Protease during Prophenoloxidase Activation. / Zhang, Rong; Cho, Hae Yun; Kim, Hyun Sic; Ma, Young Gerl; Osaki, Tsukasa; Kawabata, Shun-Ichiro; Söderhäll, Kenneth; Lee, Bok Luel.

:: Journal of Biological Chemistry, 巻 278, 番号 43, 24.10.2003, p. 42072-42079.

研究成果: ジャーナルへの寄稿記事

Zhang, Rong ; Cho, Hae Yun ; Kim, Hyun Sic ; Ma, Young Gerl ; Osaki, Tsukasa ; Kawabata, Shun-Ichiro ; Söderhäll, Kenneth ; Lee, Bok Luel. / Characterization and Properties of a 1,3-β-D-Glucan Pattern Recognition Protein of Tenebrio molitor Larvae That Is Specifically Degraded by Serine Protease during Prophenoloxidase Activation. :: Journal of Biological Chemistry. 2003 ; 巻 278, 番号 43. pp. 42072-42079.
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abstract = "Although many different pattern recognition receptors recognizing peptidoglycan and 1,3-β-D-glucan have been identified in vertebrates and insects, the molecular mechanism of these molecules in the pattern recognition and subsequent signaling is largely unknown. To gain insights into the action mechanism of 1,3-β-D-glucan pattern recognition protein in the insect prophenoloxidase (proPO) activation system, we purified a 53-kDa 1,3-β-D-glucan recognition protein (Tm-GRP) to homogeneity from the hemolymph of the mealworm, Tenebrio molitor, by using a 1,3-β-D-glucan affinity column. The purified protein specifically bound to 1,3-β-D-glucan but not to peptidoglycan. Subsequent molecular cloning revealed that Tm-GRP contains a region with close sequence similarity to bacterial glucanases. Strikingly, two catalytically important residues in glucanases are replaced with other nonhomologous amino acids in Tm-GRP. The finding suggests that Tm-GRP has evolved from an ancestral gene of glucanases but retained only the ability to recognize 1,3-β-D-glucan. A Western blot analysis of the protein level of endogenous Tm-GRP showed that the protein was specifically degraded following the activation of proPO with 1,3-β-D-glucan and calcium ion. The degradation was significantly retarded by the addition of serine protease inhibitors but not by cysteine or acidic protease inhibitor. These results suggest that 1,3-β-D-glucan pattern recognition protein is specifically degraded by serine protease(s) during proPO activation, and we propose that this degradation is an important regulatory mechanism of the activation of the proPO system.",
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T1 - Characterization and Properties of a 1,3-β-D-Glucan Pattern Recognition Protein of Tenebrio molitor Larvae That Is Specifically Degraded by Serine Protease during Prophenoloxidase Activation

AU - Zhang, Rong

AU - Cho, Hae Yun

AU - Kim, Hyun Sic

AU - Ma, Young Gerl

AU - Osaki, Tsukasa

AU - Kawabata, Shun-Ichiro

AU - Söderhäll, Kenneth

AU - Lee, Bok Luel

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N2 - Although many different pattern recognition receptors recognizing peptidoglycan and 1,3-β-D-glucan have been identified in vertebrates and insects, the molecular mechanism of these molecules in the pattern recognition and subsequent signaling is largely unknown. To gain insights into the action mechanism of 1,3-β-D-glucan pattern recognition protein in the insect prophenoloxidase (proPO) activation system, we purified a 53-kDa 1,3-β-D-glucan recognition protein (Tm-GRP) to homogeneity from the hemolymph of the mealworm, Tenebrio molitor, by using a 1,3-β-D-glucan affinity column. The purified protein specifically bound to 1,3-β-D-glucan but not to peptidoglycan. Subsequent molecular cloning revealed that Tm-GRP contains a region with close sequence similarity to bacterial glucanases. Strikingly, two catalytically important residues in glucanases are replaced with other nonhomologous amino acids in Tm-GRP. The finding suggests that Tm-GRP has evolved from an ancestral gene of glucanases but retained only the ability to recognize 1,3-β-D-glucan. A Western blot analysis of the protein level of endogenous Tm-GRP showed that the protein was specifically degraded following the activation of proPO with 1,3-β-D-glucan and calcium ion. The degradation was significantly retarded by the addition of serine protease inhibitors but not by cysteine or acidic protease inhibitor. These results suggest that 1,3-β-D-glucan pattern recognition protein is specifically degraded by serine protease(s) during proPO activation, and we propose that this degradation is an important regulatory mechanism of the activation of the proPO system.

AB - Although many different pattern recognition receptors recognizing peptidoglycan and 1,3-β-D-glucan have been identified in vertebrates and insects, the molecular mechanism of these molecules in the pattern recognition and subsequent signaling is largely unknown. To gain insights into the action mechanism of 1,3-β-D-glucan pattern recognition protein in the insect prophenoloxidase (proPO) activation system, we purified a 53-kDa 1,3-β-D-glucan recognition protein (Tm-GRP) to homogeneity from the hemolymph of the mealworm, Tenebrio molitor, by using a 1,3-β-D-glucan affinity column. The purified protein specifically bound to 1,3-β-D-glucan but not to peptidoglycan. Subsequent molecular cloning revealed that Tm-GRP contains a region with close sequence similarity to bacterial glucanases. Strikingly, two catalytically important residues in glucanases are replaced with other nonhomologous amino acids in Tm-GRP. The finding suggests that Tm-GRP has evolved from an ancestral gene of glucanases but retained only the ability to recognize 1,3-β-D-glucan. A Western blot analysis of the protein level of endogenous Tm-GRP showed that the protein was specifically degraded following the activation of proPO with 1,3-β-D-glucan and calcium ion. The degradation was significantly retarded by the addition of serine protease inhibitors but not by cysteine or acidic protease inhibitor. These results suggest that 1,3-β-D-glucan pattern recognition protein is specifically degraded by serine protease(s) during proPO activation, and we propose that this degradation is an important regulatory mechanism of the activation of the proPO system.

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