Colocalization of sterol isomerase and sigma(1) receptor at endoplasmic reticulum and nuclear envelope level.

SR31747A is a sigma ligand previously described as having original immunosuppressive properties. Two SR31747A targets were recently identified and termed sigma(1) or SR-BP-1 (SR31747A-binding protein-1) and hSI (human sterol isomerase). In order to characterize these proteins further, we examined their expression and localization at the subcellular level.

Development of a radioimmunoassay for SR 31747A, a new sigma ligand, in human plasma.

A specific and sensitive radioimmunoassay was developed for SR 31747A, a new sigma ligand, using a monoclonal antibody. This antibody was produced from spleen lymphocytes of a mouse immunized with SR 31747A coupled to bovine serum albumin via a peptide bond using SR 120684A, a succinamic acid derivative of SR 31747A. Negligible binding occurred when metabolites, obtained by chemical synthesis or by "in-vitro" incubation with hepatic microsomal fraction, were tested for cross-reactivity.

Development of a monoclonal antibody to immuno-cytochemical analysis of the cellular localization of the peripheral benzodiazepine receptor.

Based on the amino acid sequence deduced from the cloned human peripheral benzodiazepine receptor (PBR) gene, monoclonal antibody (Mab 8D7) was produced against the C-terminal fragment of the receptor. Immunoblot experiments, performed against purified PBR, indicated that the antipeptide antibody recognized, under denaturing conditions, the corresponding amino acid sequence of the PBR. When mitochondrial membranes from PBR transfected yeast or from THP1 and U937 cells were used on immunoblot analysis, a high level of immunoreactivity was observed at 18 kDa, the PBR molecular mass deduced from cDNA, establishing the specificity of the antibody for the receptor.