A protein phosphatase 2A inhibition assay for a fast and sensitive assessment of okadaic acid contamination in mussels

Toxicon. 1996 Jul;34(7):743-52. doi: 10.1016/0041-0101(96)00027-x.

Abstract

The specific inhibitory activity exerted by okadaic acid on protein phosphatase 2A was used to assess the presence of okadaic acid in mussels, using a commercially available protein phosphatase 2A preparation. Under the conditions used, okadaic acid inhibits the enzymatic activity dose-dependently, with an IC50 = 0.26 ng/ml (0.32 nM). The assay is accurate and reproducible. Okadaic acid was detected in concentrations as low as 0.063 ng/ml in aqueous solutions and 2 ng/g in mussel digestive glands. Thirty naturally contaminated mussel samples were submitted to the protein phosphatase 2A inhibition assay as well as to an ELISA assay and to a MTT cytotoxicity assay, with similar results. The proposed assay is sensitive, rapid and does not require expensive equipment. These characteristics make it a good candidate for employment in the routine assessment of okadaic acid shellfish contamination.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bivalvia / metabolism*
  • Carcinogens / analysis
  • Carcinogens / metabolism
  • Carcinogens / toxicity*
  • Coloring Agents / chemistry
  • Digestive System / metabolism
  • Dose-Response Relationship, Drug
  • Enzyme Inhibitors / metabolism
  • Enzyme Inhibitors / toxicity*
  • Enzyme-Linked Immunosorbent Assay
  • Food Contamination*
  • Lethal Dose 50
  • Okadaic Acid / analysis
  • Okadaic Acid / metabolism*
  • Okadaic Acid / toxicity
  • Phosphoprotein Phosphatases / antagonists & inhibitors*
  • Protein Phosphatase 2
  • Reproducibility of Results
  • Shellfish
  • Tetrazolium Salts / chemistry
  • Thiazoles / chemistry
  • Water Pollutants, Chemical / analysis
  • Water Pollutants, Chemical / toxicity

Substances

  • Carcinogens
  • Coloring Agents
  • Enzyme Inhibitors
  • Tetrazolium Salts
  • Thiazoles
  • Water Pollutants, Chemical
  • Okadaic Acid
  • Phosphoprotein Phosphatases
  • Protein Phosphatase 2
  • thiazolyl blue