Two cellular and subcellular locations for the peripheral-type benzodiazepine receptor in rat liver

Abstract

Determination of ligand binding properties of the peripheral benzodiazepine receptor (PBBS) in liver, in hepatocytes, and in nonparenchymal cells demonstrated the presence of receptor-specific high-affinity binding in both hepatocyte and nonhepatocyte cells. Density gradient centrifugation showed that the high-affinity receptor in hepatocytes was localised to mitochondria, whereas in nonhepatocytes it was not mitochondrial, but with a possible biliary epithelial cell plasma-membrane location. Both receptors showed the peripheraltype specific high-affinity binding of PK 11195 and Ro5 4864 and could be photolabelled as 18 kDa proteins with [ 3H]PK 14105. Immunocytochemistry showed the presence of acyl-CoA binding protein, a putative endogenous ligand for the receptor, in both cell locations. Some other properties of the PBBS were investigated in liver. Diphosphatidyl glycerol had a strong inhibitory effect on receptor binding in both liver and adrenal, with Ro5 4864 more sensitive to inhibition than PK 11195. However, whereas soybean lipid and phosphatidyl serine increased the binding of both ligands to adrenal receptor, these lipids had no effect on liver, suggesting that liver PBBS may differ from the well-characterised adrenal PBBS in some of its protein conformation. Modulators of mitochondrial respiration that also influence intermembrane contact site formation were found to elicit no marked stimulatory or inhibitory effects on PBBS ligand binding in liver, a result also found for adrenal mitochondria, suggesting that the extent of contact site formation does not influence ligand binding and that the hepatocyte receptor may not play a role in regulating mitochondrial respiration. These two cellular and subcellular locations of the PBBS in liver and the different effect of phospholipids compared to other peripheral tissues may be important for the role(s) of PBBS in liver and also for the multiple roles ascribed to the receptor and to peripheral-type benzodiazepine ligands.