Identification and characterization of leukotriene C4 and D4 receptors on a cultured smooth muscle cell line BC3H-1

N. Tamura, Devendra K. Agrawal, R. G. Townley

Research output: Contribution to journalArticle

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Abstract

We studied the characteristics of the leukotriene (LT) C4 and D4 receptors on a cultured smooth muscle cell line, BC3H-1. Specific [3H]LTC4 binding to the cell membrane was greater than 80% of total binding and saturable at a density of 3.96 ± 0.39 pmol/mg protein, with an apparent dissociation constant (Kd) of 14.3 ± 2.0 nM (n=9). The association and dissociation of [3H]LTC4 binding were rapid and apparent equilibrium conditions were established within 5 min. Calculated Kd value of [3H]LTC4 binding from the kinetic analysis was 9.9 nM. From the competition analysis, calculated Ki value of unlabeled LTC4 to compete for the specific binding of [3H]LTC4 was 9.2 nM and was in good agreement with the Kd value obtained from the Scatchard plots or kinetic analysis. The rank order of potency of the unlabeled competitors for competing specific [3H]LTC4 binding was LTC4 ≫ LTD4 > LTE4 > FPL-55712. The maximum number of binding sites (Bmax) of [3H]LTD4 in the membrane of BC3H-1 cell line was about 11 times lower than that of the [3H]LTC4. The calculated values of Kd and Bmax of [3H]LTD4 binding were 9.3 ± 0.8 nM and 0.37 ± 0.04 pmol/mg protein, respectively (n=3). The rank order of potency of the unlabeled competitors for competing specific [3H]LTD4 binding was LTD4 = LTE4 > FPL-55712 ≫ LTC4. These findings demonstrate that BC3H-1 cell line possess both LTC4 and LTD4 receptors with a predominance of LTC4 receptors. Thus BC3H-1 cell line is a good model to study the regulation of LTC4 and LTD4 receptors.

Original languageEnglish
Pages (from-to)207-216
Number of pages10
JournalLife Sciences
Volume41
Issue number2
DOIs
StatePublished - Jul 13 1987

Fingerprint

Leukotriene C4
Smooth Muscle Myocytes
Muscle
Leukotriene D4
Cells
Cell Line
Leukotriene E4
leukotriene D4 receptor
leukotriene C4 receptor
Kinetics
Cell membranes
Proteins
Binding Sites
Cell Membrane
Association reactions
Membranes

All Science Journal Classification (ASJC) codes

  • Pharmacology

Cite this

Identification and characterization of leukotriene C4 and D4 receptors on a cultured smooth muscle cell line BC3H-1. / Tamura, N.; Agrawal, Devendra K.; Townley, R. G.

In: Life Sciences, Vol. 41, No. 2, 13.07.1987, p. 207-216.

Research output: Contribution to journalArticle

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abstract = "We studied the characteristics of the leukotriene (LT) C4 and D4 receptors on a cultured smooth muscle cell line, BC3H-1. Specific [3H]LTC4 binding to the cell membrane was greater than 80{\%} of total binding and saturable at a density of 3.96 ± 0.39 pmol/mg protein, with an apparent dissociation constant (Kd) of 14.3 ± 2.0 nM (n=9). The association and dissociation of [3H]LTC4 binding were rapid and apparent equilibrium conditions were established within 5 min. Calculated Kd value of [3H]LTC4 binding from the kinetic analysis was 9.9 nM. From the competition analysis, calculated Ki value of unlabeled LTC4 to compete for the specific binding of [3H]LTC4 was 9.2 nM and was in good agreement with the Kd value obtained from the Scatchard plots or kinetic analysis. The rank order of potency of the unlabeled competitors for competing specific [3H]LTC4 binding was LTC4 ≫ LTD4 > LTE4 > FPL-55712. The maximum number of binding sites (Bmax) of [3H]LTD4 in the membrane of BC3H-1 cell line was about 11 times lower than that of the [3H]LTC4. The calculated values of Kd and Bmax of [3H]LTD4 binding were 9.3 ± 0.8 nM and 0.37 ± 0.04 pmol/mg protein, respectively (n=3). The rank order of potency of the unlabeled competitors for competing specific [3H]LTD4 binding was LTD4 = LTE4 > FPL-55712 ≫ LTC4. These findings demonstrate that BC3H-1 cell line possess both LTC4 and LTD4 receptors with a predominance of LTC4 receptors. Thus BC3H-1 cell line is a good model to study the regulation of LTC4 and LTD4 receptors.",
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N2 - We studied the characteristics of the leukotriene (LT) C4 and D4 receptors on a cultured smooth muscle cell line, BC3H-1. Specific [3H]LTC4 binding to the cell membrane was greater than 80% of total binding and saturable at a density of 3.96 ± 0.39 pmol/mg protein, with an apparent dissociation constant (Kd) of 14.3 ± 2.0 nM (n=9). The association and dissociation of [3H]LTC4 binding were rapid and apparent equilibrium conditions were established within 5 min. Calculated Kd value of [3H]LTC4 binding from the kinetic analysis was 9.9 nM. From the competition analysis, calculated Ki value of unlabeled LTC4 to compete for the specific binding of [3H]LTC4 was 9.2 nM and was in good agreement with the Kd value obtained from the Scatchard plots or kinetic analysis. The rank order of potency of the unlabeled competitors for competing specific [3H]LTC4 binding was LTC4 ≫ LTD4 > LTE4 > FPL-55712. The maximum number of binding sites (Bmax) of [3H]LTD4 in the membrane of BC3H-1 cell line was about 11 times lower than that of the [3H]LTC4. The calculated values of Kd and Bmax of [3H]LTD4 binding were 9.3 ± 0.8 nM and 0.37 ± 0.04 pmol/mg protein, respectively (n=3). The rank order of potency of the unlabeled competitors for competing specific [3H]LTD4 binding was LTD4 = LTE4 > FPL-55712 ≫ LTC4. These findings demonstrate that BC3H-1 cell line possess both LTC4 and LTD4 receptors with a predominance of LTC4 receptors. Thus BC3H-1 cell line is a good model to study the regulation of LTC4 and LTD4 receptors.

AB - We studied the characteristics of the leukotriene (LT) C4 and D4 receptors on a cultured smooth muscle cell line, BC3H-1. Specific [3H]LTC4 binding to the cell membrane was greater than 80% of total binding and saturable at a density of 3.96 ± 0.39 pmol/mg protein, with an apparent dissociation constant (Kd) of 14.3 ± 2.0 nM (n=9). The association and dissociation of [3H]LTC4 binding were rapid and apparent equilibrium conditions were established within 5 min. Calculated Kd value of [3H]LTC4 binding from the kinetic analysis was 9.9 nM. From the competition analysis, calculated Ki value of unlabeled LTC4 to compete for the specific binding of [3H]LTC4 was 9.2 nM and was in good agreement with the Kd value obtained from the Scatchard plots or kinetic analysis. The rank order of potency of the unlabeled competitors for competing specific [3H]LTC4 binding was LTC4 ≫ LTD4 > LTE4 > FPL-55712. The maximum number of binding sites (Bmax) of [3H]LTD4 in the membrane of BC3H-1 cell line was about 11 times lower than that of the [3H]LTC4. The calculated values of Kd and Bmax of [3H]LTD4 binding were 9.3 ± 0.8 nM and 0.37 ± 0.04 pmol/mg protein, respectively (n=3). The rank order of potency of the unlabeled competitors for competing specific [3H]LTD4 binding was LTD4 = LTE4 > FPL-55712 ≫ LTC4. These findings demonstrate that BC3H-1 cell line possess both LTC4 and LTD4 receptors with a predominance of LTC4 receptors. Thus BC3H-1 cell line is a good model to study the regulation of LTC4 and LTD4 receptors.

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