The concentrations of total protein, mucus glycoprotein, cyclic-AMP, and apolipoproteins A-I, A-II, and B were determined in the gallbladder bile of patients with cholesterol gallbladder stones and in stone-free controls. The total protein content was significantly increased in gallstone patients (2.03 +/- 0.6 versus 1.31 +/- 0.67 mg/ml; p less than 0.05), as was the mucus glycoprotein concentration (380 +/- 88.5 versus 128 +/- 57.2 micrograms/ml; p less than 0.05). The cyclic-AMP concentration in the gallbladder fluid was increased up to 91 +/- 20 pmol/100 microliters in the gallstone subjects, as compared with 46 +/- 26 pmol/100 microliters (p less than 0.01) in stone-free controls. Cyclic-AMP concentrations correlated positively with the glycoprotein content of the bile in cholesterol gallstone patients (r = 0.66; p less than 0.05). The apolipoprotein concentrations were determined by the radial immundiffusion technique. The corresponding values for patients with stones and controls were 7.5 +/- 0.8 versus 3.0 +/- 0.8 for Apo A-I (p less than 0.025), 10.4 +/- 0.6 versus 6.3 +/- 1.3 for Apo A-II (p less than 0.02), and 1.9 +/- 0.5 versus 1.6 +/- 0.2 mg/dl for Apo B (NS), respectively. Biliary proteins probably play an important role in the nucleation process during the pathogenesis of cholesterol gallbladder stones.