Molecular and ligand-binding characterization of the σ-receptor in the Jurkat human T lymphocyte cell line

Malliga E. Ganapathy, Puttur D Prasad, Wei Huang, Pankaj Seth, Frederick H. Leibach, Vadivel Ganapathy

Research output: Contribution to journalArticle

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Abstract

The σ binding site present in the Jurkat human T lymphocyte cell line was investigated. Jurkat cell membranes were found to have a single saturable binding site for [3H]haloperidol, a ligand (dissociation constant, 3.9 ± 0.3 nM). The binding of [3H]haloperidol was inhibited by several σ ligands. Northern analysis and reverse transcription-polymerase chain reaction provided evidence for the expression of the recently cloned type 1 σ- receptor (σ-R1) in Jurkat cells. The σ-R1 cDNA cloned from these cells was functional in heterologous expression systems. When expressed in mammalian celts, the cDNA-induced binding was saturable with dissociation constants of 1.9 ± 0.3 nM for [3H]haloperidol and 12 ± 2 nM for (+)- pentazocine. The binding of [3H]progesterone, a putative endogenous ligand to σ-R1, to the Jurkat cell σ-receptor could be directly demonstrated by using heterologously expressed σ-R1 cDNA. The binding of [3H]progesterone was saturable, with a dissociation constant of 88 ± 7 nM. Progesterone and haloperidol interacted with the receptor competitively. Reverse transcription-polymerase chain reaction also produced evidence for the existence of an alternatively spliced σ-R1 variant in Jurkat cells. This splice variant was found to be nonfunctional in ligand binding assays. This constitutes the first report on the molecular characterization of the σ- receptor in immune cells.

Original languageEnglish (US)
Pages (from-to)251-260
Number of pages10
JournalJournal of Pharmacology and Experimental Therapeutics
Volume289
Issue number1
StatePublished - Apr 1999

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Jurkat Cells
Haloperidol
Ligands
Progesterone
T-Lymphocytes
Cell Line
Complementary DNA
Reverse Transcription
Binding Sites
Pentazocine
Polymerase Chain Reaction
Cell Membrane

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology

Cite this

Molecular and ligand-binding characterization of the σ-receptor in the Jurkat human T lymphocyte cell line. / Ganapathy, Malliga E.; Prasad, Puttur D; Huang, Wei; Seth, Pankaj; Leibach, Frederick H.; Ganapathy, Vadivel.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 289, No. 1, 04.1999, p. 251-260.

Research output: Contribution to journalArticle

Ganapathy, Malliga E. ; Prasad, Puttur D ; Huang, Wei ; Seth, Pankaj ; Leibach, Frederick H. ; Ganapathy, Vadivel. / Molecular and ligand-binding characterization of the σ-receptor in the Jurkat human T lymphocyte cell line. In: Journal of Pharmacology and Experimental Therapeutics. 1999 ; Vol. 289, No. 1. pp. 251-260.
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