LIQUID CHROMATOGRAPHY-LINKED PROTEIN PHOSPHATASE BIOASSAY - A HIGHLY SENSITIVE MARINE BIOSCREEN FOR OKADAIC ACID AND RELATED DIARRHETIC SHELLFISH TOXINS
LIQUID CHROMATOGRAPHY-LINKED PROTEIN PHOSPHATASE BIOASSAY - A HIGHLY SENSITIVE MARINE BIOSCREEN FOR OKADAIC ACID AND RELATED DIARRHETIC SHELLFISH TOXINS
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DOI：
10.1016/0041-0101(91)90021-i
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发表时间：
1991-01-01
期刊：
影响因子：
2.8
通讯作者：
HOLMES, CFB
中科院分区：
文献类型：
--
作者：
HOLMES, CFB
Okadaic acid and dinophysistoxin-1 were resolved by liquid chromatography, then identified and quantitated by specific inhibition of both protein phosphatase-1 and -2A (PP1/PP2A) catalytic subunits in a P-32-phosphorylase a phosphatase radioassay. Based on the IC50 for PP2A inhibition (0.2 nM), the procedure has a detection sensitivity of less than 10 pg okadaic acid. Confirmative identification by PP1 inhibition (IC50 = 19 nM) requires 500 pg okadaic acid. Analyses of methanolic extracts from control, "okadaic acid spiked" and suspected diarrhetic mussels showed the bioscreen to be accurate, reproducible and identified okadaic acid/dinophysistoxin-1 in Canadian shellfish for the first time. In addition, a protein phosphatase inhibitor distinct from okadaic acid/dinophysistoxin-1 was identified in diarrhetic mussels with a potency equivalent to 900 ng okadaic acid/g digestive tract. Protein phosphatase inhibition probably underlies the biological activity of okadaic acid as a diarrhetic shellfish toxin and tumour promoter (COHEN, P., HOLMES, C. F. B. and TSUKITANI, Y. (1990), TIBS 15, 98-102). The liquid chromatography-linked protein phosphatase bioscreen should therefore facilitate identification of novel toxins comprising diarrhetic profiles in infested shellfish.
