Microcystin‐LR Detected in a Low Molecular Weight  Fraction from a Crude Extract of Zoanthus sociatus

Abstract

Cnidarian constitutes a great source of bioactive compounds. However, research involving peptides from organisms belonging to the order Zoanthidea has received very little attention, contrasting to the numerous studies of the order Actiniaria, from which hundreds of toxic peptides and proteins have been reported. In this work, we performed a mass spectrometry analysis of a low molecular weight (LMW) fraction previously reported as lethal to mice. The low molecular weight (LMW) fraction was obtained by gel filtration of a Zoanthus sociatus (order Zoanthidea) crude extract with a Sephadex G-50, and then analyzed by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI-TOF/TOF) mass spectrometry (MS) in positive ion reflector mode from m/z 700 to m/z 4000. Afterwards, some of the most intense and representative MS ions were fragmented by MS/MS with no significant results obtained by Protein Pilot protein identification software and the Mascot algorithm search. However, microcystin masses were detected by mass-matching against libraries of non-ribosomal peptide database (NORINE). Subsequent reversed-phase C18 HPLC (in isocratic elution mode) and mass spectrometry analyses corroborated the presence of the cyanotoxin Microcystin-LR (MC-LR). To the best of our knowledge, this finding constitutes the first report of MC-LR in Z. sociatus, and one of the few evidences of such cyanotoxin in cnidarians.

In Vitro Toxicity Evaluation of Cyanotoxins Cylindrospermopsin and Microcystin-LR on Human Kidney HEK293 Cells

“…The LC-MS/MS was performed in a nano-LC coupled to a hybrid Ion trap mass spectrometer (LTQ Orbitrap Velos Pro-ETD) as described by Campos et al [ 78 ] and Dominguez-Pérez et al [ 79 ]. Peptides were separated by reverse phase chromatography on a 20 mm × 100 µm C18 precolumn followed by a 100 mm × 75 µm C18 column (particle size 5 µm, NanoSeparations, Nieuwkoop, The Netherlands) in a linear gradient of acetonitrile (2% to 95% v / v ) in FA (0.1% v / v ), at a flow rate of 0.3 µL/min (total elution time 70 min).…”

Coelenterate Toxins, Its Pharmaceutical and Therapeutic Effects

“…Acid-sensing ion channels (ASICs) are H+-gated Na+channels that belongs toENaC/degenerin superfamily of sodium channels. ASICs possess numerous functions such as sensory perception, synaptic plasticity, learning, memory formation, cell migration and proliferation, nociception, and neurodegenerative disorders [42] (fig. 2).…”

Physiological and Metabolic Responses of Marine Mussels Exposed to Toxic Cyanobacteria Microcystis aeruginosa and Chrysosporum ovalisporum

“…The LC-MS/MS was carried out as described previously in a nano-LC coupled to a hybrid Ion trap mass spectrometer (LTQ Orbitrap Velos Pro-ETD, Thermo Scientific, Waltham, MA, USA) [32,68]. Peptides were separated by reverse phase chromatography on a 20 mm × 100 µm C18 precolumn followed by a 100 mm × 75 µm C18 column (particle size 5 µm, NanoSeparations, Nieuwkoop, Netherlands) in a linear gradient of acetonitrile (2% to 95% v/v) in FA (0.1% v/v), at a flow rate of 0.3 µL/min (total elution time 70 min).…”