Photoaffinity labeling of a P3 purinoceptor-like protein purified from rat brain membranes

Biochem Biophys Res Commun. 1996 Feb 15;219(2):469-74. doi: 10.1006/bbrc.1996.0257.

Abstract

We have found a novel adenosine binding protein in rat brain membranes. Based on the ligand binding specificities, this protein is not classified into any known P1 or P2 purinoceptors but is likely to be classified into the putative P3 purinoceptor which was recently proposed. This P3 purinoceptor-like protein was partially purified from rat brain membranes and characterized further by photoaffinity labeling with 8-azidoadenosine-5'-[gamma-32P]triphosphate. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the labeled preparations revealed that the probe is incorporated specifically into a single band with an approximate molecular size of 54 kDa.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenine Nucleotides / metabolism*
  • Adenine Nucleotides / pharmacology
  • Adenosine / analogs & derivatives
  • Adenosine / metabolism*
  • Adenosine / pharmacology
  • Adenosine Triphosphate / analogs & derivatives*
  • Adenosine Triphosphate / metabolism
  • Adenosine-5'-(N-ethylcarboxamide)
  • Affinity Labels / metabolism
  • Animals
  • Azides / metabolism*
  • Brain / metabolism*
  • Cell Fractionation
  • Cell Membrane / metabolism
  • Chromatography
  • Chromatography, Affinity
  • Durapatite
  • Electrophoresis, Polyacrylamide Gel
  • Kinetics
  • Ligands
  • Molecular Weight
  • Rats
  • Receptors, Purinergic / isolation & purification
  • Receptors, Purinergic / metabolism*
  • Ribonucleotides / metabolism
  • Ribonucleotides / pharmacology

Substances

  • Adenine Nucleotides
  • Affinity Labels
  • Azides
  • Ligands
  • Receptors, Purinergic
  • Ribonucleotides
  • Adenosine-5'-(N-ethylcarboxamide)
  • 8-azidoadenosine 5'-triphosphate
  • Adenosine Triphosphate
  • Durapatite
  • Adenosine