Cross-linking of neuropeptide Y to its receptor on rat brain membranes.

Academic Article

Abstract

  • The receptor for neuropeptide Y (NPY) was identified on rat brain membranes after covalent labeling with 125I-NPY using the homobifunctional cross-linkers disuccinimido suberate and disuccinimido dithiobis(propionate) and the heterobifunctional photoactive cross-linker succinimido 4-azidobenzoate. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by autoradiography revealed the presence of two bands at Mr 62,000 and 39,000. Both species showed the same high affinity for 125I-NPY. Exposure to reducing agents did not change the migration of these bands. When the NPY receptor complex was solubilized from the membranes with 1% Triton X-100 and analyzed by gel filtration chromatography, it eluted from a Fractogel TSK 55F column as a peak at approximately 65 kDa. This peak was asymmetric with a shoulder of radioactivity that probably reflects the smaller receptor species. These data indicate that the NPY receptor on rat brain membranes is a monomeric 58-kDa unit (62 kDa minus the mass of the cross-linked NPY) without covalently or noncovalently linked subunits. The smaller 39-kDa species may be an immature form of the 62-kDa species, a second distinct receptor, or a degradation product of the 62-kDa band.
  • Published In

    Keywords

  • Animals, Brain, Cross-Linking Reagents, Kinetics, Male, Membranes, Molecular Weight, Neuropeptide Y, Rats, Rats, Inbred Strains, Receptors, Neuropeptide Y, Receptors, Neurotransmitter
  • Digital Object Identifier (doi)

    Author List

  • Mannon PJ; Taylor IL; Kaiser LM; Nguyen TD
  • Start Page

  • G637
  • End Page

  • G643
  • Volume

  • 256
  • Issue

  • 3 Pt 1