Kinin B1 receptors on rabbit aorta smooth muscle cells in culture were investigated. [3H]Des-Arg10-kallidin labeled a single site in cells at early passage with an equilibrium dissociation constant of 258 pM and a maximal binding density of approximately 680 sites/cell. Treatment of the same cells for 18 h with epidermal growth factor increased the binding density over 6-fold without affecting the ligand's affinity. At latter passages, the density of binding sites was found to increase and the growth factor had a much less pronounced effect. The rank order of potencies for agonist inhibition of binding (des-Arg10-kallidin > des-Arg9-BK = kallidin > bradykinin) was consistent with the specific labeling of a B1 receptor. Also, [3H]des-Arg10-kallidin binding was potently inhibited by the B1 receptor antagonist des-Arg9[Leu8]bradykinin but not by the B2 receptor antagonist Hoe 140. The agonists were found to stimulate phosphoinositide hydrolysis in the smooth muscle cells with an order of potencies that reflected their binding assay activities. Des-Arg9[Leu8] BK blocked the des-Arg10-kallidin response with a potency consistent with its known B1 receptor activity while Hoe 140 was inactive. These results demonstrate the presence of inducible B1 receptors on rabbit aorta smooth muscle cells in culture that couple to phospholipase C activation. These cells should be useful in future studies of the mechanisms and factors involved in the regulation of expression of the B1 receptor.