Purification and characterization of neurotensin receptor from rat brain with special reference to comparison between newborn and adult age rats

Peptides. 1991 Sep-Oct;12(5):1001-6. doi: 10.1016/0196-9781(91)90050-y.

Abstract

Scatchard analysis of saturation curves was performed to compared newborn and adult rat neurotensin receptor using [3H] neurotensin as a tracer. The membrane fraction of newborn rat cerebral cortex has a single population of neurotensin receptor (Kd = 0.13 nM, Bmax = 710 fmol/mg protein), whereas adults have two distinct neurotensin binding sites (high affinity site, Kd1 = 0.13 nM; low affinity site, Kd2 = 20 nM). High affinity neurotensin receptor, solubilized with digitonin, was purified from newborn rat cortex by affinity chromatography. An overall purification of 14,000-fold was achieved. The binding of [3H] neurotensin to the purified receptor is saturable and specific, with a Kd of 0.45 nM. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the presence of 2-mercaptoethanol revealed purified material of a single major band of Mr = 55,000.

MeSH terms

  • Aging
  • Animals
  • Animals, Newborn
  • Brain / growth & development*
  • Brain / metabolism*
  • Cell Membrane / metabolism
  • Cerebral Cortex / growth & development
  • Cerebral Cortex / metabolism
  • Chromatography, Affinity
  • Electrophoresis, Polyacrylamide Gel
  • Hypothalamus / growth & development
  • Hypothalamus / metabolism
  • Kinetics
  • Molecular Weight
  • Neurotensin / metabolism*
  • Rats
  • Rats, Inbred Strains
  • Receptors, Neurotensin
  • Receptors, Neurotransmitter / isolation & purification
  • Receptors, Neurotransmitter / metabolism*

Substances

  • Receptors, Neurotensin
  • Receptors, Neurotransmitter
  • Neurotensin