Zeta (ζ), the opioid growth factor receptor

identification and characterization of binding subunits

Ian S. Zagon, Steven Goodman, Patricia J. McLaughlin

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

The zeta (ζ) opioid receptor mediates the activity of the opioid growth factor, [Met5]-enkephalin, a peptide that regulates developmental events in a variety of normal and tumorigenic tissues and cells, including the nervous system. To identify the binding subunit(s) of the ζ receptor, protein blots of rat cerebellar proteins from 6-day-old animals were separated by sodium dodecyl sulfate 10% polyacrylamide gel electrophoresis (SDS-PAGE) and electrophoretically transferred onto nitrocellulose. Ligand blotting of these blots with 1.5 nM[125I]-[Met5]-enkephalin revealed four binding polypeptides of 32, 30, 17, and 16 kDa. Binding occurred at concentrations relevant to the Kd of the receptor, was blocked by cold ligand and opioid antagonists, and exhibited a stereospecific response. No binding was recorded in the adult rat cerebellum. Subcellular fractionation studies using ligand blotting and receptor-binding analysis indicated that these binding subunits were associated with the nucleus. Two-dimensional protein analysis using non-equilibrium pH gradient electrophoresis (NEPHGE) SDS-PAGE of 6-day-old cerebellum and ligand blotting showed that the 32-, 30-, 17-, and 16-kDa subunits have basic isoelectric points. Two-dimensional chymotryptic peptide mapping showed a strong homology between the 32- and 30-kDa subunits, but not with the 17- and 16-kDa polypeptides. The 17- and 16-kDa subunits had only a partial homology to each other. These results are consistent with the biology, biochemistry, and pharmacology of the ζ receptor, and are the first to identify and characterize the binding polypeptides of an opioid receptor that has important growth-related properties.

Original languageEnglish (US)
Pages (from-to)50-56
Number of pages7
JournalBrain Research
Volume605
Issue number1
DOIs
StatePublished - Mar 5 1993
Externally publishedYes

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Ligands
Peptides
Enkephalins
Opioid Receptors
Sodium Dodecyl Sulfate
Cerebellum
Polyacrylamide Gel Electrophoresis
Proteins
Proton-Motive Force
Peptide Mapping
Collodion
Narcotic Antagonists
Isoelectric Point
Biochemistry
Opioid Analgesics
Nervous System
Electrophoresis
Intercellular Signaling Peptides and Proteins
Pharmacology
methionine-enkephalin receptor

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)
  • Molecular Biology
  • Clinical Neurology
  • Developmental Biology

Cite this

Zeta (ζ), the opioid growth factor receptor : identification and characterization of binding subunits. / Zagon, Ian S.; Goodman, Steven; McLaughlin, Patricia J.

In: Brain Research, Vol. 605, No. 1, 05.03.1993, p. 50-56.

Research output: Contribution to journalArticle

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