Summary
A method is described for measuring rapid, specific, and saturable binding of the skin irritant and tumour-promoting secretagogue thapsigargin (sesquiterpene lactone) to the microsomal fraction from mouse brain. Employing the tritium-labelled compound its apparent dissociation constant,K d, and the maximal amount of bindingB max are shown to be 9.8 nM and 1.9 pmol/mg protein respectively. Such aK d for thapsigargin is similar to (a) its IC50 value for inhibiting Ca2+ uptake in the microsomal fraction from rat brain and (b) its EC50 values for inducing a rise in the cytoplasmic Ca2+ concentration of human platelets and histamine release from rat peritoneal mast cells. A positive correlation is found between the binding affinities of thapsigargin, thapsitranstagin, and trilobolide, their potencies as secretagogues and their lipophilicities. This correlation does not extend to the skin-irritant activities of the compounds thus emphasizing that their mechanism of action is unlike that of 12-O-tetradecanoylphorbol 13-acetate.
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Abbreviations
- Tg:
-
thapsigargin
- Tt:
-
thapsitranstagin
- Tr:
-
trilobolide
- TPA:
-
12-O-tetradecanoylphorbol 13-acetate
- K i,K d :
-
apparent inhibition and dissociation constants
- ID 550 :
-
irritant dose for 50% response after 5 h
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For II see Hergenhahn et al. 1991
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Christensen, S.B., Hergenhahn, M., Roeser, H. et al. Toxicodynamics of tumour promoters of mouse skin III. Specific binding of the tumour promoter thapsigargin as measured by the cold-acetone filter assay. J Cancer Res Clin Oncol 118, 344–348 (1992). https://doi.org/10.1007/BF01294438
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DOI: https://doi.org/10.1007/BF01294438