Structural insights to the heterotetrameric interaction between the vibrio parahaemolyticus pirAvp and pirBvp 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

Research output: Contribution to journalArticle

1 Citation (Scopus)

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 PirAvp and PirBvp 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 PirAvp and PirBvp has not yet been determined. Here, we used isothermal titration calorimetry (ITC) to measure the binding affinity of PirAvp and PirBvp. Since the dissociation constant (Kd = 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 PirAvp-PirBvp interaction, first by using gel filtration to evaluate the molecular weight of the PirAvp/PirBvp complex, and then by using cross-linking and hydrogen-deuterium exchange (HDX) mass spectrometry to further understand the interaction interface between PirAvp and PirBvp. 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 PirBvp N-terminal pore-forming domain and should be helpful for devising effective anti-AHPND strategies in the future.

Original languageEnglish
Article number233
JournalToxins
Volume11
Issue number4
DOIs
Publication statusPublished - 2019 Apr

Fingerprint

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

All Science Journal Classification (ASJC) codes

  • Toxicology
  • Health, Toxicology and Mutagenesis

Cite this

@article{6343a669373948c98e26887eb479773f,
title = "Structural insights to the heterotetrameric interaction between the vibrio parahaemolyticus pirAvp and pirBvp 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 PirAvp and PirBvp 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 PirAvp and PirBvp has not yet been determined. Here, we used isothermal titration calorimetry (ITC) to measure the binding affinity of PirAvp and PirBvp. Since the dissociation constant (Kd = 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 PirAvp-PirBvp interaction, first by using gel filtration to evaluate the molecular weight of the PirAvp/PirBvp complex, and then by using cross-linking and hydrogen-deuterium exchange (HDX) mass spectrometry to further understand the interaction interface between PirAvp and PirBvp. 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 PirBvp N-terminal pore-forming domain and should be helpful for devising effective anti-AHPND strategies in the future.",
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",
month = "4",
doi = "10.3390/toxins11040233",
language = "English",
volume = "11",
journal = "Toxins",
issn = "2072-6651",
publisher = "Multidisciplinary Digital Publishing Institute (MDPI)",
number = "4",

}

Structural insights to the heterotetrameric interaction between the vibrio parahaemolyticus pirAvp and pirBvp 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.

In: Toxins, Vol. 11, No. 4, 233, 04.2019.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Structural insights to the heterotetrameric interaction between the vibrio parahaemolyticus pirAvp and pirBvp 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

Y1 - 2019/4

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 PirAvp and PirBvp 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 PirAvp and PirBvp has not yet been determined. Here, we used isothermal titration calorimetry (ITC) to measure the binding affinity of PirAvp and PirBvp. Since the dissociation constant (Kd = 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 PirAvp-PirBvp interaction, first by using gel filtration to evaluate the molecular weight of the PirAvp/PirBvp complex, and then by using cross-linking and hydrogen-deuterium exchange (HDX) mass spectrometry to further understand the interaction interface between PirAvp and PirBvp. 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 PirBvp 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 PirAvp and PirBvp 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 PirAvp and PirBvp has not yet been determined. Here, we used isothermal titration calorimetry (ITC) to measure the binding affinity of PirAvp and PirBvp. Since the dissociation constant (Kd = 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 PirAvp-PirBvp interaction, first by using gel filtration to evaluate the molecular weight of the PirAvp/PirBvp complex, and then by using cross-linking and hydrogen-deuterium exchange (HDX) mass spectrometry to further understand the interaction interface between PirAvp and PirBvp. 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 PirBvp N-terminal pore-forming domain and should be helpful for devising effective anti-AHPND strategies in the future.

UR - http://www.scopus.com/inward/record.url?scp=85065317672&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85065317672&partnerID=8YFLogxK

U2 - 10.3390/toxins11040233

DO - 10.3390/toxins11040233

M3 - Article

C2 - 31013623

AN - SCOPUS:85065317672

VL - 11

JO - Toxins

JF - Toxins

SN - 2072-6651

IS - 4

M1 - 233

ER -