Eicosanoids are thought to play an important role in the pathogenesis of inflammatory skin diseases. The object of the present study was the detection and characterization of putative 12(S)-hydroxyeicosatetraenoic acid (12(S)-HETE) binding sites in normal human keratinocytes. Keratinocytes were obtained from foreskin and dermatome-shaved normal human skin. Radioligand binding assays were performed with 12(S)-[3H]HETE on cultured cells. Analysis of saturation curves suggested a one-site model for 12(S)-HETE binding with a KD of 3.84 +/- 0.18 nM and receptor number Bmax of 2.32 +/- 0.12 x 10(5) per cell. Ligand binding was reversible. The rank order of potency in competition for 12(S)-[3H]HETE was 12(S)-HETE > 12(R)- HETE > or = leukotriene B4. Preincubation of cells with 12(S)-HETE (2 x 10(-6) M) resulted in down-regulation of the binding site by approximately 50%. The identification and characterization of specific 12(S)-HETE binding sites on normal human keratinocytes should enable further elucidation of the role of 12-HETE in cutaneous biology and in the pathophysiology of psoriasis and other inflammatory and hyperproliferative dermatoses.