A human cancer cell line (COLO 16) derived originally from an epidermal squamous cell carcinoma was found to possess adenylate cyclase responsiveness to beta-adrenergic agonists. 2. The adenylate cyclase response was characterized with respect to activation constants (KA) for various beta-adrenergic agonists and inhibition constants (Ki) for antagonists. 3. Intact cells responded with dose-dependent increases in production of cyclic adenosine 3':5'-monophosphate. 4. Properties of the beta-adrenergic receptor were evaluated by using the specific binding of [3H]propranolol to cell membranes. Specific binding was saturable, with KD 5.79 nmol/l and binding sites 0.68 pmol/mg of protein. 5. Competition for binding to cell membranes was shown by beta-adrenergic agonists and antagonists and was stereospecific. There was close agreement between the affinity of these various agents on adenylate cyclase and receptor binding. 6. It is likely that the beta-adrenergic receptor-linked adenylate cyclase in COLO 16 cells represents persistence in a cancer cell line of a receptor present normally in epidermal cells.
