Purification by affinity chromatography of the glycine receptor of rat spinal cord

J Biol Chem. 1982 Aug 25;257(16):9389-93.


The glycine receptor of rat spinal cord was solubilized with the nonionic detergent Triton X-100 and subsequently purified by affinity chromatography on aminostrychnine-agarose and wheat germ agglutinin-Sepharose. An overall purification of 1950-fold was achieved. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and mercaptoethanol revealed three glycine receptor-associated polypeptides of Mr = 48,000, 58,000, and 93,000. [3H]Strychnine was incorporated irreversibly into the Mr = 48,000 polypeptide upon UV-illumination. The dissociation constant (KD) of [3H]strychnine binding to the purified glycine receptor was 9.3 +/- 0.6 nM. The glycine receptor agonists glycine, beta-alanine, and taurine inhibited the binding of [3H]strychnine to the purified receptor. Gel filtration and sedimentation in sucrose/H2O and sucrose/D2O gradients gave a Stokes radius of 7.7 nm, a partial specific volume of 0.780 +/- 0.005 ml/g and a sedimentation coefficient s20,w of 8.2 +/- 0.2 S for the purified glycine receptor. From these data, a molecular weight of 246,000 +/- 6,000 was calculated for the glycine receptor protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Affinity Labels
  • Animals
  • Centrifugation, Density Gradient
  • Chromatography, Affinity / methods*
  • Electrophoresis, Polyacrylamide Gel
  • Molecular Weight
  • Rats
  • Rats, Inbred Strains
  • Receptors, Cell Surface / isolation & purification*
  • Receptors, Cell Surface / metabolism
  • Receptors, Glycine
  • Spinal Cord / analysis*
  • Strychnine / metabolism


  • Affinity Labels
  • Receptors, Cell Surface
  • Receptors, Glycine
  • Strychnine