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Structural and functional study of reconstituted peripheral benzodiazepine receptor
J J Lacapère1, F Delavoie, H Li
1Unité INSERM U410, Faculté de Médecine Xavier Bichat, 16 rue Henri Huchard, Paris Cedex 18, 75870, France.
Biochemical and Biophysical Research Communications
|June 8, 2001
Summary
Recombinant mouse peripheral-type benzodiazepine receptor (PBR) protein was successfully reconstituted into liposomes. The purified PBR monomer demonstrated binding to drug ligands and cholesterol.
Area of Science:
- Biochemistry
- Molecular Biology
- Structural Biology
Background:
- The peripheral-type benzodiazepine receptor (PBR) plays a crucial role in various cellular processes.
- Understanding the functional unit of PBR is essential for drug development and cellular studies.
Purpose of the Study:
- To overexpress and purify recombinant mouse PBR protein.
- To reconstitute the purified PBR into liposomes and characterize its functional properties.
Main Methods:
- Recombinant PBR overexpression in E. coli and purification using His. Bind resin.
- Liposome reincorporation using Bio-Beads SM2 for detergent removal.
- Electron microscopy (negative staining, freeze-fracture) and ligand binding assays.
Main Results:
- Proteoliposomes containing recombinant PBR were successfully formed.
- Electron microscopy revealed transmembranous particles consistent with PBR monomers.
- Reconstituted PBR bound PK 11195 and cholesterol with nanomolar affinity.
Conclusions:
- A PBR monomer represents the minimal functional unit of the receptor.
- The reconstituted PBR retains its ability to bind both drug ligands and cholesterol.
- This study provides insights into the structural and functional characteristics of PBR.