Maitotoxin-4, a Novel MTX Analog Produced by Gambierdiscus excentricus

Maitotoxins (MTXs) are among the most potent toxins known. These toxins are produced by epi-benthic dinoflagellates of the genera Gambierdiscus and Fukuyoa and may play a role in causing the symptoms associated with Ciguatera Fish Poisoning. A recent survey revealed that, of the species tested, the...

Descripción completa

Detalles Bibliográficos
Autores principales: Pisapia, Francesco, Sibat, Manoëlla, Herrenknecht, Christine, Lhaute, Korian, Gaiani, Greta, Ferron, Pierre-Jean, Fessard, Valérie, Fraga, Santiago, Nascimento, Silvia M., Litaker, R. Wayne, Holland, William C., Roullier, Catherine, Hess, Philipp
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5532662/
https://www.ncbi.nlm.nih.gov/pubmed/28696398
http://dx.doi.org/10.3390/md15070220
_version_ 1783253505076625408
author Pisapia, Francesco
Sibat, Manoëlla
Herrenknecht, Christine
Lhaute, Korian
Gaiani, Greta
Ferron, Pierre-Jean
Fessard, Valérie
Fraga, Santiago
Nascimento, Silvia M.
Litaker, R. Wayne
Holland, William C.
Roullier, Catherine
Hess, Philipp
author_facet Pisapia, Francesco
Sibat, Manoëlla
Herrenknecht, Christine
Lhaute, Korian
Gaiani, Greta
Ferron, Pierre-Jean
Fessard, Valérie
Fraga, Santiago
Nascimento, Silvia M.
Litaker, R. Wayne
Holland, William C.
Roullier, Catherine
Hess, Philipp
author_sort Pisapia, Francesco
collection PubMed
description Maitotoxins (MTXs) are among the most potent toxins known. These toxins are produced by epi-benthic dinoflagellates of the genera Gambierdiscus and Fukuyoa and may play a role in causing the symptoms associated with Ciguatera Fish Poisoning. A recent survey revealed that, of the species tested, the newly described species from the Canary Islands, G. excentricus, is one of the most maitotoxic. The goal of the present study was to characterize MTX-related compounds produced by this species. Initially, lysates of cells from two Canary Island G. excentricus strains VGO791 and VGO792 were partially purified by (i) liquid-liquid partitioning between dichloromethane and aqueous methanol followed by (ii) size-exclusion chromatography. Fractions from chromatographic separation were screened for MTX toxicity using both the neuroblastoma neuro-2a (N2a) cytotoxicity and Ca(2+) flux functional assays. Fractions containing MTX activity were analyzed using liquid chromatography coupled to high-resolution mass spectrometry (LC-HRMS) to pinpoint potential MTX analogs. Subsequent non-targeted HRMS analysis permitted the identification of a novel MTX analog, maitotoxin-4 (MTX4, accurate mono-isotopic mass of 3292.4860 Da, as free acid form) in the most toxic fractions. HRMS/MS spectra of MTX4 as well as of MTX are presented. In addition, crude methanolic extracts of five other strains of G. excentricus and 37 other strains representing one Fukuyoa species and ten species, one ribotype and one undetermined strain/species of Gambierdiscus were screened for the presence of MTXs using low resolution tandem mass spectrometry (LRMS/MS). This targeted analysis indicated the original maitotoxin (MTX) was only present in one strain (G. australes S080911_1). Putative maitotoxin-2 (p-MTX2) and maitotoxin-3 (p-MTX3) were identified in several other species, but confirmation was not possible because of the lack of reference material. Maitotoxin-4 was detected in all seven strains of G. excentricus examined, independently of their origin (Brazil, Canary Islands and Caribbean), and not detected in any other species. MTX4 may therefore serve as a biomarker for the highly toxic G. excentricus in the Atlantic area.
format Online
Article
Text
id pubmed-5532662
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-55326622017-08-09 Maitotoxin-4, a Novel MTX Analog Produced by Gambierdiscus excentricus Pisapia, Francesco Sibat, Manoëlla Herrenknecht, Christine Lhaute, Korian Gaiani, Greta Ferron, Pierre-Jean Fessard, Valérie Fraga, Santiago Nascimento, Silvia M. Litaker, R. Wayne Holland, William C. Roullier, Catherine Hess, Philipp Mar Drugs Article Maitotoxins (MTXs) are among the most potent toxins known. These toxins are produced by epi-benthic dinoflagellates of the genera Gambierdiscus and Fukuyoa and may play a role in causing the symptoms associated with Ciguatera Fish Poisoning. A recent survey revealed that, of the species tested, the newly described species from the Canary Islands, G. excentricus, is one of the most maitotoxic. The goal of the present study was to characterize MTX-related compounds produced by this species. Initially, lysates of cells from two Canary Island G. excentricus strains VGO791 and VGO792 were partially purified by (i) liquid-liquid partitioning between dichloromethane and aqueous methanol followed by (ii) size-exclusion chromatography. Fractions from chromatographic separation were screened for MTX toxicity using both the neuroblastoma neuro-2a (N2a) cytotoxicity and Ca(2+) flux functional assays. Fractions containing MTX activity were analyzed using liquid chromatography coupled to high-resolution mass spectrometry (LC-HRMS) to pinpoint potential MTX analogs. Subsequent non-targeted HRMS analysis permitted the identification of a novel MTX analog, maitotoxin-4 (MTX4, accurate mono-isotopic mass of 3292.4860 Da, as free acid form) in the most toxic fractions. HRMS/MS spectra of MTX4 as well as of MTX are presented. In addition, crude methanolic extracts of five other strains of G. excentricus and 37 other strains representing one Fukuyoa species and ten species, one ribotype and one undetermined strain/species of Gambierdiscus were screened for the presence of MTXs using low resolution tandem mass spectrometry (LRMS/MS). This targeted analysis indicated the original maitotoxin (MTX) was only present in one strain (G. australes S080911_1). Putative maitotoxin-2 (p-MTX2) and maitotoxin-3 (p-MTX3) were identified in several other species, but confirmation was not possible because of the lack of reference material. Maitotoxin-4 was detected in all seven strains of G. excentricus examined, independently of their origin (Brazil, Canary Islands and Caribbean), and not detected in any other species. MTX4 may therefore serve as a biomarker for the highly toxic G. excentricus in the Atlantic area. MDPI 2017-07-11 /pmc/articles/PMC5532662/ /pubmed/28696398 http://dx.doi.org/10.3390/md15070220 Text en © 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Pisapia, Francesco
Sibat, Manoëlla
Herrenknecht, Christine
Lhaute, Korian
Gaiani, Greta
Ferron, Pierre-Jean
Fessard, Valérie
Fraga, Santiago
Nascimento, Silvia M.
Litaker, R. Wayne
Holland, William C.
Roullier, Catherine
Hess, Philipp
Maitotoxin-4, a Novel MTX Analog Produced by Gambierdiscus excentricus
title Maitotoxin-4, a Novel MTX Analog Produced by Gambierdiscus excentricus
title_full Maitotoxin-4, a Novel MTX Analog Produced by Gambierdiscus excentricus
title_fullStr Maitotoxin-4, a Novel MTX Analog Produced by Gambierdiscus excentricus
title_full_unstemmed Maitotoxin-4, a Novel MTX Analog Produced by Gambierdiscus excentricus
title_short Maitotoxin-4, a Novel MTX Analog Produced by Gambierdiscus excentricus
title_sort maitotoxin-4, a novel mtx analog produced by gambierdiscus excentricus
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5532662/
https://www.ncbi.nlm.nih.gov/pubmed/28696398
http://dx.doi.org/10.3390/md15070220
work_keys_str_mv AT pisapiafrancesco maitotoxin4anovelmtxanalogproducedbygambierdiscusexcentricus
AT sibatmanoella maitotoxin4anovelmtxanalogproducedbygambierdiscusexcentricus
AT herrenknechtchristine maitotoxin4anovelmtxanalogproducedbygambierdiscusexcentricus
AT lhautekorian maitotoxin4anovelmtxanalogproducedbygambierdiscusexcentricus
AT gaianigreta maitotoxin4anovelmtxanalogproducedbygambierdiscusexcentricus
AT ferronpierrejean maitotoxin4anovelmtxanalogproducedbygambierdiscusexcentricus
AT fessardvalerie maitotoxin4anovelmtxanalogproducedbygambierdiscusexcentricus
AT fragasantiago maitotoxin4anovelmtxanalogproducedbygambierdiscusexcentricus
AT nascimentosilviam maitotoxin4anovelmtxanalogproducedbygambierdiscusexcentricus
AT litakerrwayne maitotoxin4anovelmtxanalogproducedbygambierdiscusexcentricus
AT hollandwilliamc maitotoxin4anovelmtxanalogproducedbygambierdiscusexcentricus
AT roulliercatherine maitotoxin4anovelmtxanalogproducedbygambierdiscusexcentricus
AT hessphilipp maitotoxin4anovelmtxanalogproducedbygambierdiscusexcentricus