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Toxins 2010, 2(1), 195-204; doi:10.3390/toxins2010195
Article

PP2A Inhibition Assay Using Recombinant Enzyme for Rapid Detection of Okadaic Acid and Its Analogs in Shellfish

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Received: 27 November 2009; in revised form: 18 January 2010 / Accepted: 19 January 2010 / Published: 25 January 2010
(This article belongs to the Special Issue Marine Biotoxins: Novel Issues about Old Compounds)
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Abstract: Okadaic acid and its analogs (OAs) responsible for diarrhetic shellfish poisoning (DSP) strongly inhibit protein phosphatase 2A (PP2A) and thus are quantifiable by measuring the extent of the enzyme inhibition. In this study, we evaluated the suitability of the catalytic subunit of recombinant human PP2A (rhPP2Ac) for use in a microplate OA assay. OA, dinophysistoxin-1(DTX1), and hydrolyzate of 7-O-palmitoyl-OA strongly inhibited rhPP2Ac activity with IC50 values of 0.095, 0.104, and 0.135 nM, respectively. The limits of detection and quantitation for OA in the digestive gland of scallops and mussels were 0.0348 μg/g and 0.0611 μg/g respectively, and, when converted to the whole meat basis, are well below the regulation level proposed by EU (0.16 μg/g whole meat). A good correlation with LC-MS data was demonstrated, the correlation coefficient being 0.996 with the regression slope of 1.097.
Keywords: diarrhetic shellfish poisoning (DSP); okadaic acid (OA); protein phosphatase 2A (PP2A) inhibition assay; digestive gland (DG) diarrhetic shellfish poisoning (DSP); okadaic acid (OA); protein phosphatase 2A (PP2A) inhibition assay; digestive gland (DG)
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Ikehara, T.; Imamura, S.; Yoshino, A.; Yasumoto, T. PP2A Inhibition Assay Using Recombinant Enzyme for Rapid Detection of Okadaic Acid and Its Analogs in Shellfish. Toxins 2010, 2, 195-204.

AMA Style

Ikehara T, Imamura S, Yoshino A, Yasumoto T. PP2A Inhibition Assay Using Recombinant Enzyme for Rapid Detection of Okadaic Acid and Its Analogs in Shellfish. Toxins. 2010; 2(1):195-204.

Chicago/Turabian Style

Ikehara, Tsuyoshi; Imamura, Shihoko; Yoshino, Atsushi; Yasumoto, Takeshi. 2010. "PP2A Inhibition Assay Using Recombinant Enzyme for Rapid Detection of Okadaic Acid and Its Analogs in Shellfish." Toxins 2, no. 1: 195-204.



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