Identification of a 27-kDa high affinity phenylalkylamine-binding polypeptide as the σ1 binding site by photoaffinity labeling and ligand-directed antibodies

Fabian F. Moebius, Gregory G. Burrows, Markus Hanner, Eduard Schmid, Jörg Striessnig, Hartmut Glossmann

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

The verapamil-like arylazide (-)-[3H]azidopamil specifically photoaffinity labeled two low molecular mass polypeptides, with apparent molecular masses of 22 and 27 KDa, in the endoplasmic reticulum of guinea pig liver, kidney, adrenal gland, and lung. It was recently shown that the 22-kDa polypeptide binds the anti-ischemic phenylalkylamine (-)-[3H]emopamil and other anti-ischemic drugs with high affinity. We now provide evidence that the photolabeling of the 27-kDa polypeptide is blocked by nanomolar concentrations of σ ligands [order of potency, haloperidol > pentazocine > 1,3-ditolylguanidine > dextromethorphan > (+)-SKF10,047]. The apparent affinities of these and other drugs closely corresponded to those for 1,3-[3H]ditolylguanidine-labeled σ binding sites. Based on its high affinity for the (+)-enantiomer [but not the (-)-enantiomer] of SKF10.047 (Ki = 51 nM), pentazocine (Ki = 3 nM), and dextromethorphan (Ki = 30 nM), the (-)-[3H]azidopamil-labeled site on the 27-kDa polypeptide was classified as being of the σ1 subtype. Using antiphenylalkylamine antibodies, we developed a novel immunological detection method that allows the rapid and sensitive staining of the photolabeled 27-kDa polypeptide after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. We conclude that the phenylalkylamines emopamil and azidopamil represent a novel class of σ ligands, highly suitable for the further structural characterization of polypeptides carrying σ1 binding sites.

Original languageEnglish (US)
Pages (from-to)966-971
Number of pages6
JournalMolecular Pharmacology
Volume44
Issue number5
StatePublished - Nov 1993
Externally publishedYes

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Binding Sites
Ligands
Peptides
Antibodies
Dextromethorphan
Pentazocine
Haloperidol
Adrenal Glands
Verapamil
Sodium Dodecyl Sulfate
Endoplasmic Reticulum
Pharmaceutical Preparations
Polyacrylamide Gel Electrophoresis
Guinea Pigs
Staining and Labeling
Kidney
Lung
Liver
LU 49888
1,3-ditolylguanidine

ASJC Scopus subject areas

  • Pharmacology

Cite this

Identification of a 27-kDa high affinity phenylalkylamine-binding polypeptide as the σ1 binding site by photoaffinity labeling and ligand-directed antibodies. / Moebius, Fabian F.; Burrows, Gregory G.; Hanner, Markus; Schmid, Eduard; Striessnig, Jörg; Glossmann, Hartmut.

In: Molecular Pharmacology, Vol. 44, No. 5, 11.1993, p. 966-971.

Research output: Contribution to journalArticle

Moebius, Fabian F. ; Burrows, Gregory G. ; Hanner, Markus ; Schmid, Eduard ; Striessnig, Jörg ; Glossmann, Hartmut. / Identification of a 27-kDa high affinity phenylalkylamine-binding polypeptide as the σ1 binding site by photoaffinity labeling and ligand-directed antibodies. In: Molecular Pharmacology. 1993 ; Vol. 44, No. 5. pp. 966-971.
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