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

Research output: Contribution to journalArticle

9 Citations (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 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.

Original languageEnglish
Article number233
JournalToxins
Volume11
Issue number4
DOIs
Publication statusPublished - Apr 1 2019

Keywords

  • AHPND
  • PirA
  • PirB
  • Protein-protein interaction
  • Vibrio parahaemolyticus

ASJC Scopus subject areas

  • Toxicology
  • Health, Toxicology and Mutagenesis

Fingerprint Dive into the research topics of 'Structural insights to the heterotetrameric interaction between the vibrio parahaemolyticus pirA <sup>vp</sup> and pirB <sup>vp</sup> toxins and activation of the cry-like pore-forming domain'. Together they form a unique fingerprint.

  • Cite this