Abstract
Palytoxin (PLTX), produced by dinoflagellates from the genus Ostreopsis was first discovered, isolated, and purified from zoanthids belonging to the genus Palythoa. The detection of this toxin in contaminated shellfish is essential for human health preservation. A broad range of studies indicate that mammalian Na+,K+-ATPase is a high affinity cellular receptor for PLTX. The toxin converts the pump into an open channel that stimulates sodium influx and potassium efflux. In this work we develop a detection method for PLTX based on its binding to the Na+,K+-ATPase. The method was developed by using the phenomenon of surface plasmon resonance (SPR) to monitor biomolecular reactions. This technique does not require any labeling of components. The interaction of PLTX over immobilized Na+,K+-ATPase is quantified by injecting different concentrations of toxin in the biosensor and checking the binding rate constant (kobs). From the representation of kobs versus PLTX concentration, the kinetic equilibrium dissociation constant (KD) for the PLTX-Na+,K+-ATPase association can be calculated. The value of this constant is KD = 6.38 × 10−7 ± 6.67 × 10−8 M PLTX. In this way the PLTX-Na+,K+-ATPase association was used as a suitable method for determination of the toxin concentration in a sample. This method represents a new and useful approach to easily detect the presence of PLTX-like compounds in marine products using the mechanism of action of these toxins and in this way reduce the use of other more expensive and animal based methods.
Highlights
IntroductionPalytoxin (PLTX), isolated from the marine soft coral (genus Palythoa), is one of the most poisonous non-protein substances known to date
Palytoxin (PLTX), isolated from the marine soft coral, is one of the most poisonous non-protein substances known to date
Na+,K+-ATPase was used as the ligand attached to the sensor surface and PLTX in solution was used as the ligate
Summary
Palytoxin (PLTX), isolated from the marine soft coral (genus Palythoa), is one of the most poisonous non-protein substances known to date. It is common in tropical and subtropical waters and may accumulate at very high levels in fish and crabs [1]. The large spatial expansion of this toxin has led to intensification of research towards optimizations of methods for determination of PLTX presence and toxicity. This toxin is a large, very complex molecule with both lipophilic and hydrophilic areas, and has the longest chain of continuous carbon atoms known to exit in a natural product. Several molecules related with PLTX have been described: palytoxin-b, homopalytoxin, bishomopalytoxin, neopalytoxin, deoxypalytoxin, 42-hydroxypalytoxin, ostreocin-d, ovatoxin-a, -b,
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