Surface Plasmon Resonance Biosensor Method for Palytoxin Detection Based on Na+,K+-ATPase Affinity
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 tha...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2013-12-01
|
| Series: | Toxins |
| Subjects: | |
| Online Access: | http://www.mdpi.com/2072-6651/6/1/96 |
| _version_ | 1818038707691716608 |
|---|---|
| author | Amparo Alfonso María-José Pazos Andrea Fernández-Araujo Araceli Tobio Carmen Alfonso Mercedes R. Vieytes Luis M. Botana |
| author_facet | Amparo Alfonso María-José Pazos Andrea Fernández-Araujo Araceli Tobio Carmen Alfonso Mercedes R. Vieytes Luis M. Botana |
| author_sort | Amparo Alfonso |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-10T07:47:01Z |
| format | Article |
| id | doaj.art-cbc9cec92db94826947acdcf3b3e59c9 |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-12-10T07:47:01Z |
| publishDate | 2013-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-cbc9cec92db94826947acdcf3b3e59c92022-12-22T01:57:10ZengMDPI AGToxins2072-66512013-12-01619610710.3390/toxins6010096toxins6010096Surface Plasmon Resonance Biosensor Method for Palytoxin Detection Based on Na+,K+-ATPase AffinityAmparo Alfonso0María-José Pazos1Andrea Fernández-Araujo2Araceli Tobio3Carmen Alfonso4Mercedes R. Vieytes5Luis M. Botana6Department of Pharmacology, Veterinary School, University of Santiago de Compostela, 27002 Lugo, SpainConfocal and Electronic Microscopy Unit, RIAIDT, University of Santiago de Compostela, 27002 Lugo, SpainDepartment of Pharmacology, Veterinary School, University of Santiago de Compostela, 27002 Lugo, SpainDepartment of Pharmacology, Veterinary School, University of Santiago de Compostela, 27002 Lugo, SpainCifga Laboratory, Santo Domingo Square, 20, 5ª, 27001 Lugo, SpainDepartment of Physiology, Veterinary School, University of Santiago de Compostela, 27002 Lugo, SpainDepartment of Pharmacology, Veterinary School, University of Santiago de Compostela, 27002 Lugo, SpainPalytoxin (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.http://www.mdpi.com/2072-6651/6/1/96palytoxinNa+,K+-ATPasesurface plasmon resonance biosensorOstreopsis siamensis |
| spellingShingle | Amparo Alfonso María-José Pazos Andrea Fernández-Araujo Araceli Tobio Carmen Alfonso Mercedes R. Vieytes Luis M. Botana Surface Plasmon Resonance Biosensor Method for Palytoxin Detection Based on Na+,K+-ATPase Affinity Toxins palytoxin Na+,K+-ATPase surface plasmon resonance biosensor Ostreopsis siamensis |
| title | Surface Plasmon Resonance Biosensor Method for Palytoxin Detection Based on Na+,K+-ATPase Affinity |
| title_full | Surface Plasmon Resonance Biosensor Method for Palytoxin Detection Based on Na+,K+-ATPase Affinity |
| title_fullStr | Surface Plasmon Resonance Biosensor Method for Palytoxin Detection Based on Na+,K+-ATPase Affinity |
| title_full_unstemmed | Surface Plasmon Resonance Biosensor Method for Palytoxin Detection Based on Na+,K+-ATPase Affinity |
| title_short | Surface Plasmon Resonance Biosensor Method for Palytoxin Detection Based on Na+,K+-ATPase Affinity |
| title_sort | surface plasmon resonance biosensor method for palytoxin detection based on na k atpase affinity |
| topic | palytoxin Na+,K+-ATPase surface plasmon resonance biosensor Ostreopsis siamensis |
| url | http://www.mdpi.com/2072-6651/6/1/96 |
| work_keys_str_mv | AT amparoalfonso surfaceplasmonresonancebiosensormethodforpalytoxindetectionbasedonnakatpaseaffinity AT mariajosepazos surfaceplasmonresonancebiosensormethodforpalytoxindetectionbasedonnakatpaseaffinity AT andreafernandezaraujo surfaceplasmonresonancebiosensormethodforpalytoxindetectionbasedonnakatpaseaffinity AT aracelitobio surfaceplasmonresonancebiosensormethodforpalytoxindetectionbasedonnakatpaseaffinity AT carmenalfonso surfaceplasmonresonancebiosensormethodforpalytoxindetectionbasedonnakatpaseaffinity AT mercedesrvieytes surfaceplasmonresonancebiosensormethodforpalytoxindetectionbasedonnakatpaseaffinity AT luismbotana surfaceplasmonresonancebiosensormethodforpalytoxindetectionbasedonnakatpaseaffinity |