Cloning and expression of a pharmacologically unique bovine peripheral-type benzodiazepine receptor isoquinoline binding protein

Anthony L. Parola, Donald G. Stump, David J. Pepperl, Karl E. Krueger, John W Regan, Hugh E. Laird

Research output: Contribution to journalArticle

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Abstract

High affinity binding of isoquinolines, such as PK 11195, is a conserved feature of peripheral-type benzodiazepine receptors (PER) across species. However, species differences in PBR ligand binding have been described based on the affinity for N1-alkyl-1,4-benzodiazepines, such as Ro5-4864. Ro5-4864 binds with high affinity to the rat receptor but has low affinity for the bovine PBR. Photolabeling with an isoquinoline ligand, [3H]PK 14105, identifies a 17-kDa protein, the PBR isoquinoline binding protein (PBR/IBP), in both species. To further elucidate the role of the PBR/IBP in determining PBR benzodiazepine and isoquinoline binding characteristics, the bovine PBR/IBP was cloned and expressed. Using a cDNA encoding a rat PBR/IBP to screen a fetal bovine adrenal cDNA library, a bovine cDNA encoding a polypeptide of 169 residues was cloned. The bovine and rat PBR/IBPs had similar hydropathy profiles exhibiting five potential transmembrane domains. Transfecting the cloned bovine PBR/IBP cDNA into COS-7 cells resulted in an 11-fold increase in the density of high affinity [3H]PK 11195 binding sites which had only low affinity for Ro5-4864. Expression of the bovine PBR/IBP yields a receptor which is pharmacologically distinct from both endogenous COS-7 PBR and the rat PBR based on the affinity for several N1-alkyl-1,4-benzodiazepine ligands. These results suggest the PBR/IBP is the minimal functional component required for PBR ligand binding characteristics and the different protein sequences account for the species differences in PBR benzodiazepine ligand binding.

Original languageEnglish (US)
Pages (from-to)14082-14087
Number of pages6
JournalJournal of Biological Chemistry
Volume266
Issue number21
StatePublished - 1991

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Cloning
GABA-A Receptors
Organism Cloning
Carrier Proteins
Rats
Ligands
Complementary DNA
Benzodiazepines
Isoquinolines
isoquinoline
COS Cells
Gene Library
Membrane Potentials
Proteins
Binding Sites
Peptides

ASJC Scopus subject areas

  • Biochemistry

Cite this

Cloning and expression of a pharmacologically unique bovine peripheral-type benzodiazepine receptor isoquinoline binding protein. / Parola, Anthony L.; Stump, Donald G.; Pepperl, David J.; Krueger, Karl E.; Regan, John W; Laird, Hugh E.

In: Journal of Biological Chemistry, Vol. 266, No. 21, 1991, p. 14082-14087.

Research output: Contribution to journalArticle

Parola, Anthony L. ; Stump, Donald G. ; Pepperl, David J. ; Krueger, Karl E. ; Regan, John W ; Laird, Hugh E. / Cloning and expression of a pharmacologically unique bovine peripheral-type benzodiazepine receptor isoquinoline binding protein. In: Journal of Biological Chemistry. 1991 ; Vol. 266, No. 21. pp. 14082-14087.
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