Structural insights to the heterotetrameric interaction between the vibrio parahaemolyticus pirA vp and pirB vp toxins and activation of the cry-like pore-forming domain

Shin Jen Lin, Yi Fan Chen, Kai Cheng Hsu, Yun Ling Chen, Tzu Ping Ko, Chu Fang Lo, Han Ching Wang, Hao Ching Wang

研究成果: 雜誌貢獻文章

1 引文 (Scopus)

摘要

Acute hepatopancreatic necrosis disease (AHPND) is a newly emergent penaeid shrimp disease which can cause 70-100% mortality in Penaeus vannamei and Penaeus monodon, and has resulted in enormous economic losses since its appearance. AHPND is caused by the specific strains of Vibrio parahaemolyticus that harbor the pVA1 plasmid and express PirA vp and PirB vp toxins. These two toxins have been reported to form a binary complex. When both are present, they lead to the death of shrimp epithelial cells in the hepatopancreas and cause the typical histological symptoms of AHPND. However, the binding mode of PirA vp and PirB vp has not yet been determined. Here, we used isothermal titration calorimetry (ITC) to measure the binding affinity of PirA vp and PirB vp . Since the dissociation constant (K d = 7.33 ± 1.20 μM) was considered too low to form a sufficiently stable complex for X-ray crystallographic analysis, we used alternative methods to investigate PirA vp -PirB vp interaction, first by using gel filtration to evaluate the molecular weight of the PirA vp /PirB vp complex, and then by using cross-linking and hydrogen-deuterium exchange (HDX) mass spectrometry to further understand the interaction interface between PirA vp and PirB vp . Based on these results, we propose a heterotetrameric interaction model of this binary toxin complex. This model provides insight of how conformational changes might activate the PirB vp N-terminal pore-forming domain and should be helpful for devising effective anti-AHPND strategies in the future.
原文英語
文章編號233
期刊Toxins
11
發行號4
DOIs
出版狀態已發佈 - 四月 1 2019

指紋

Vibrio parahaemolyticus
Penaeidae
Chemical activation
Necrosis
Hepatopancreas
Calorimetry
Deuterium
Ports and harbors
Titration
Gel Chromatography
Mass spectrometry
Hydrogen
Mass Spectrometry
Plasmids
Molecular Weight
Gels
Epithelial Cells
Molecular weight
Economics
X-Rays

ASJC Scopus subject areas

  • Toxicology
  • Health, Toxicology and Mutagenesis

引用此文

Structural insights to the heterotetrameric interaction between the vibrio parahaemolyticus pirA vp and pirB vp toxins and activation of the cry-like pore-forming domain. / Lin, Shin Jen; Chen, Yi Fan; Hsu, Kai Cheng; Chen, Yun Ling; Ko, Tzu Ping; Lo, Chu Fang; Wang, Han Ching; Wang, Hao Ching.

於: Toxins, 卷 11, 編號 4, 233, 01.04.2019.

研究成果: 雜誌貢獻文章

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title = "Structural insights to the heterotetrameric interaction between the vibrio parahaemolyticus pirA vp and pirB vp toxins and activation of the cry-like pore-forming domain",
abstract = "Acute hepatopancreatic necrosis disease (AHPND) is a newly emergent penaeid shrimp disease which can cause 70-100{\%} mortality in Penaeus vannamei and Penaeus monodon, and has resulted in enormous economic losses since its appearance. AHPND is caused by the specific strains of Vibrio parahaemolyticus that harbor the pVA1 plasmid and express PirA vp and PirB vp toxins. These two toxins have been reported to form a binary complex. When both are present, they lead to the death of shrimp epithelial cells in the hepatopancreas and cause the typical histological symptoms of AHPND. However, the binding mode of PirA vp and PirB vp has not yet been determined. Here, we used isothermal titration calorimetry (ITC) to measure the binding affinity of PirA vp and PirB vp . Since the dissociation constant (K d = 7.33 ± 1.20 μM) was considered too low to form a sufficiently stable complex for X-ray crystallographic analysis, we used alternative methods to investigate PirA vp -PirB vp interaction, first by using gel filtration to evaluate the molecular weight of the PirA vp /PirB vp complex, and then by using cross-linking and hydrogen-deuterium exchange (HDX) mass spectrometry to further understand the interaction interface between PirA vp and PirB vp . Based on these results, we propose a heterotetrameric interaction model of this binary toxin complex. This model provides insight of how conformational changes might activate the PirB vp N-terminal pore-forming domain and should be helpful for devising effective anti-AHPND strategies in the future.",
keywords = "AHPND, PirA, PirB, Protein-protein interaction, Vibrio parahaemolyticus",
author = "Lin, {Shin Jen} and Chen, {Yi Fan} and Hsu, {Kai Cheng} and Chen, {Yun Ling} and Ko, {Tzu Ping} and Lo, {Chu Fang} and Wang, {Han Ching} and Wang, {Hao Ching}",
year = "2019",
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T1 - Structural insights to the heterotetrameric interaction between the vibrio parahaemolyticus pirA vp and pirB vp toxins and activation of the cry-like pore-forming domain

AU - Lin, Shin Jen

AU - Chen, Yi Fan

AU - Hsu, Kai Cheng

AU - Chen, Yun Ling

AU - Ko, Tzu Ping

AU - Lo, Chu Fang

AU - Wang, Han Ching

AU - Wang, Hao Ching

PY - 2019/4/1

Y1 - 2019/4/1

N2 - Acute hepatopancreatic necrosis disease (AHPND) is a newly emergent penaeid shrimp disease which can cause 70-100% mortality in Penaeus vannamei and Penaeus monodon, and has resulted in enormous economic losses since its appearance. AHPND is caused by the specific strains of Vibrio parahaemolyticus that harbor the pVA1 plasmid and express PirA vp and PirB vp toxins. These two toxins have been reported to form a binary complex. When both are present, they lead to the death of shrimp epithelial cells in the hepatopancreas and cause the typical histological symptoms of AHPND. However, the binding mode of PirA vp and PirB vp has not yet been determined. Here, we used isothermal titration calorimetry (ITC) to measure the binding affinity of PirA vp and PirB vp . Since the dissociation constant (K d = 7.33 ± 1.20 μM) was considered too low to form a sufficiently stable complex for X-ray crystallographic analysis, we used alternative methods to investigate PirA vp -PirB vp interaction, first by using gel filtration to evaluate the molecular weight of the PirA vp /PirB vp complex, and then by using cross-linking and hydrogen-deuterium exchange (HDX) mass spectrometry to further understand the interaction interface between PirA vp and PirB vp . Based on these results, we propose a heterotetrameric interaction model of this binary toxin complex. This model provides insight of how conformational changes might activate the PirB vp N-terminal pore-forming domain and should be helpful for devising effective anti-AHPND strategies in the future.

AB - Acute hepatopancreatic necrosis disease (AHPND) is a newly emergent penaeid shrimp disease which can cause 70-100% mortality in Penaeus vannamei and Penaeus monodon, and has resulted in enormous economic losses since its appearance. AHPND is caused by the specific strains of Vibrio parahaemolyticus that harbor the pVA1 plasmid and express PirA vp and PirB vp toxins. These two toxins have been reported to form a binary complex. When both are present, they lead to the death of shrimp epithelial cells in the hepatopancreas and cause the typical histological symptoms of AHPND. However, the binding mode of PirA vp and PirB vp has not yet been determined. Here, we used isothermal titration calorimetry (ITC) to measure the binding affinity of PirA vp and PirB vp . Since the dissociation constant (K d = 7.33 ± 1.20 μM) was considered too low to form a sufficiently stable complex for X-ray crystallographic analysis, we used alternative methods to investigate PirA vp -PirB vp interaction, first by using gel filtration to evaluate the molecular weight of the PirA vp /PirB vp complex, and then by using cross-linking and hydrogen-deuterium exchange (HDX) mass spectrometry to further understand the interaction interface between PirA vp and PirB vp . Based on these results, we propose a heterotetrameric interaction model of this binary toxin complex. This model provides insight of how conformational changes might activate the PirB vp N-terminal pore-forming domain and should be helpful for devising effective anti-AHPND strategies in the future.

KW - AHPND

KW - PirA

KW - PirB

KW - Protein-protein interaction

KW - Vibrio parahaemolyticus

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DO - 10.3390/toxins11040233

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