Leukotriene C4 ([3H] - LTC4) binding to membranes isolated from a hamster smooth muscle cell line (DDTIMF2) |
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Authors: | Mike A Clark Michael Cook Seymour Mong G Kurt Hogaboom Robert Shorr Jeffrey Stadel Stanley T Crooke |
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Institution: | Smith Kline and French Laboratories 1500 Spring Garden Street Philadelphia, PA 19101, USA |
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Abstract: | Leukotriene C4 (LTC4) has been demonstrated to induce contraction of the smooth muscle cell line DDTIMF2. A partially purified membrane fraction obtained from these cells exhibited a high affinity binding site for LTC4. Binding of 3H]-LTC4 was saturable, specific and reversible with a dissociation constant (Kd) of 21 ± 4 nM. The maximum number of binding sites (Bmax) was 55 ± 5 pmol/mg of protein. Specificity was demonstrated in competition studies in which the Ki of LTC4 against specifically bound 3H] - LTC4 was 12 nM whereas Leukotriene D4 (LTD4) and Leukotriene E4 (LTE4) had a Ki of 38 ± 4 and 4.7 ± 0.5 nM respectively. A previously described antagonist of leukotriene-induced smooth muscle contraction PFL 55712 had a Ki of 23 ± 2 nM as determined by competition binding experiments. |
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