Abstract

The binding of [3H]leukotriene (LT) C4 to membranes from human bronchi has been characterized. The specific binding, measured at 4 degrees C, was very rapid, equilibrium being attained within 1 to 5 min. The binding was also rapidly reversible and saturable (maximum binding = 187 pmol/mg of protein). LTC4, LTD4, LTE4 and the LT antagonist FPL 55712 competed with [3H]LTC4 for its binding sites, with the following IC50 values: 0.12; 2.3; 30; and 20 microM. Therefore, the binding sites displayed a higher affinity for LTC4 than for the other sulfidopeptide LTs. Computer-assisted analysis of either the saturation or the competition curves for LTC4 indicated the existence of two classes of binding sites with different affinities (Kd1 and Kd2 = 70 nM and 0.58 microM, respectively), in agreement with the curved semilog plot of the dissociation time course. CaCl2 or MgCl2 increased and GTP or 5'-guanylylimidodiphosphate did not decrease the specific binding. In addition, the distribution of the binding sites for [3H]LTC4 along the human respiratory tree was investigated. At a fixed (10 nM) [3H]LTC4 concentration, membranes obtained from bronchi removed at different levels of the airway tree did not bind LTC4 in a significantly different amount. This is compatible with the finding that LTC4 receptors should be present on bronchi of various caliber, as both small and large airways respond to LTs.