14(R),15(S)-epoxyeicosatrienoic acid (14(R),15(S)-EET) receptor in guinea pig mononuclear cell membranes

P. Y K Wong, K. T. Lin, Y. T. Yan, D. Ahern, J. Iles, S. Y. Shen, R. K. Bhatt, J R Falck

Research output: Contribution to journalArticle

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Abstract

A high affinity binding site for 14(R),15(S)-EET, one of the major cytochrome P-450 metabolites of arachidonic acid (AA) in blood vessels, liver, kidney and urine of patients with pregnancy-induced hyertension, has been identified in a membrane preparation from guinea pig mononuclear (GPM) cells. Using a radioligand assay, binding of 14(R),15(S)-[3H]EET to its receptor site was saturable, specific and reversible. Scatchard analysis of saturation binding studies yielded a dissociation constant (K(d)) of 5.7 X 10-9 M, and maximum number of binding sites (B(max)) of 2.4 pmol/mg membrane protein. The specificity of the binding site was determined by competition studies. 14(S),15(R)-EET and 8,9-EET had a K(i) of 6.3 and 8.8 nM, respectively, followed by 12(R)-HETE and LTD4. 12(S)-HETE and 5,6-EET were even less effective as a competitive inhibitor of radioligand binding with K(i) values from 2 to 20 μM. Receptor antagonists for TxA2, LTB4, LTD4 and PAF failed to displace 14(R),15(S)-[3H]EET from its binding site on GPM cell membranes. The results correlate well with the reported biological functions of 14,15-EET. In view of its potent biological activities, 14,15-EET may exert its cellular function through the binding and activation of its stereo-specific cell surface binding sites or receptor.

Original languageEnglish (US)
Pages (from-to)199-208
Number of pages10
JournalJournal of Lipid Mediators
Volume6
Issue number1-3
StatePublished - 1992

Fingerprint

Cell membranes
Guinea Pigs
Binding Sites
Cell Membrane
Acids
12-Hydroxy-5,8,10,14-eicosatetraenoic Acid
Leukotriene D4
Radioligand Assay
Leukotriene B4
Blood vessels
Cell Surface Receptors
Metabolites
Bioactivity
Arachidonic Acid
Liver
Cytochrome P-450 Enzyme System
Blood Vessels
Assays
Membrane Proteins
Chemical activation

Keywords

  • 14,15-EET stereoisomers
  • 14,15-epoxyeicosatrienoic acid receptor
  • guinea pig mononuclear cell membrane

ASJC Scopus subject areas

  • Immunology
  • Pharmacology

Cite this

Wong, P. Y. K., Lin, K. T., Yan, Y. T., Ahern, D., Iles, J., Shen, S. Y., ... Falck, J. R. (1992). 14(R),15(S)-epoxyeicosatrienoic acid (14(R),15(S)-EET) receptor in guinea pig mononuclear cell membranes. Journal of Lipid Mediators, 6(1-3), 199-208.

14(R),15(S)-epoxyeicosatrienoic acid (14(R),15(S)-EET) receptor in guinea pig mononuclear cell membranes. / Wong, P. Y K; Lin, K. T.; Yan, Y. T.; Ahern, D.; Iles, J.; Shen, S. Y.; Bhatt, R. K.; Falck, J R.

In: Journal of Lipid Mediators, Vol. 6, No. 1-3, 1992, p. 199-208.

Research output: Contribution to journalArticle

Wong, PYK, Lin, KT, Yan, YT, Ahern, D, Iles, J, Shen, SY, Bhatt, RK & Falck, JR 1992, '14(R),15(S)-epoxyeicosatrienoic acid (14(R),15(S)-EET) receptor in guinea pig mononuclear cell membranes', Journal of Lipid Mediators, vol. 6, no. 1-3, pp. 199-208.
Wong, P. Y K ; Lin, K. T. ; Yan, Y. T. ; Ahern, D. ; Iles, J. ; Shen, S. Y. ; Bhatt, R. K. ; Falck, J R. / 14(R),15(S)-epoxyeicosatrienoic acid (14(R),15(S)-EET) receptor in guinea pig mononuclear cell membranes. In: Journal of Lipid Mediators. 1992 ; Vol. 6, No. 1-3. pp. 199-208.
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abstract = "A high affinity binding site for 14(R),15(S)-EET, one of the major cytochrome P-450 metabolites of arachidonic acid (AA) in blood vessels, liver, kidney and urine of patients with pregnancy-induced hyertension, has been identified in a membrane preparation from guinea pig mononuclear (GPM) cells. Using a radioligand assay, binding of 14(R),15(S)-[3H]EET to its receptor site was saturable, specific and reversible. Scatchard analysis of saturation binding studies yielded a dissociation constant (K(d)) of 5.7 X 10-9 M, and maximum number of binding sites (B(max)) of 2.4 pmol/mg membrane protein. The specificity of the binding site was determined by competition studies. 14(S),15(R)-EET and 8,9-EET had a K(i) of 6.3 and 8.8 nM, respectively, followed by 12(R)-HETE and LTD4. 12(S)-HETE and 5,6-EET were even less effective as a competitive inhibitor of radioligand binding with K(i) values from 2 to 20 μM. Receptor antagonists for TxA2, LTB4, LTD4 and PAF failed to displace 14(R),15(S)-[3H]EET from its binding site on GPM cell membranes. The results correlate well with the reported biological functions of 14,15-EET. In view of its potent biological activities, 14,15-EET may exert its cellular function through the binding and activation of its stereo-specific cell surface binding sites or receptor.",
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AU - Wong, P. Y K

AU - Lin, K. T.

AU - Yan, Y. T.

AU - Ahern, D.

AU - Iles, J.

AU - Shen, S. Y.

AU - Bhatt, R. K.

AU - Falck, J R

PY - 1992

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N2 - A high affinity binding site for 14(R),15(S)-EET, one of the major cytochrome P-450 metabolites of arachidonic acid (AA) in blood vessels, liver, kidney and urine of patients with pregnancy-induced hyertension, has been identified in a membrane preparation from guinea pig mononuclear (GPM) cells. Using a radioligand assay, binding of 14(R),15(S)-[3H]EET to its receptor site was saturable, specific and reversible. Scatchard analysis of saturation binding studies yielded a dissociation constant (K(d)) of 5.7 X 10-9 M, and maximum number of binding sites (B(max)) of 2.4 pmol/mg membrane protein. The specificity of the binding site was determined by competition studies. 14(S),15(R)-EET and 8,9-EET had a K(i) of 6.3 and 8.8 nM, respectively, followed by 12(R)-HETE and LTD4. 12(S)-HETE and 5,6-EET were even less effective as a competitive inhibitor of radioligand binding with K(i) values from 2 to 20 μM. Receptor antagonists for TxA2, LTB4, LTD4 and PAF failed to displace 14(R),15(S)-[3H]EET from its binding site on GPM cell membranes. The results correlate well with the reported biological functions of 14,15-EET. In view of its potent biological activities, 14,15-EET may exert its cellular function through the binding and activation of its stereo-specific cell surface binding sites or receptor.

AB - A high affinity binding site for 14(R),15(S)-EET, one of the major cytochrome P-450 metabolites of arachidonic acid (AA) in blood vessels, liver, kidney and urine of patients with pregnancy-induced hyertension, has been identified in a membrane preparation from guinea pig mononuclear (GPM) cells. Using a radioligand assay, binding of 14(R),15(S)-[3H]EET to its receptor site was saturable, specific and reversible. Scatchard analysis of saturation binding studies yielded a dissociation constant (K(d)) of 5.7 X 10-9 M, and maximum number of binding sites (B(max)) of 2.4 pmol/mg membrane protein. The specificity of the binding site was determined by competition studies. 14(S),15(R)-EET and 8,9-EET had a K(i) of 6.3 and 8.8 nM, respectively, followed by 12(R)-HETE and LTD4. 12(S)-HETE and 5,6-EET were even less effective as a competitive inhibitor of radioligand binding with K(i) values from 2 to 20 μM. Receptor antagonists for TxA2, LTB4, LTD4 and PAF failed to displace 14(R),15(S)-[3H]EET from its binding site on GPM cell membranes. The results correlate well with the reported biological functions of 14,15-EET. In view of its potent biological activities, 14,15-EET may exert its cellular function through the binding and activation of its stereo-specific cell surface binding sites or receptor.

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