Cross-linking of atrial natriuretic peptide to binding sites in rat olfactory bulb membranes
Binding sites for /sup 125/I-atrial natriuretic peptide (ANP)2 in rat olfactory bulb membranes have been studied using pharmacological and biochemical methods. Various unlabeled ANP-related peptides were tested for the ability to inhibit the binding of the radioligand in membrane binding assays. ANP(92-126) and ANP(99-126) were the most potent inhibitors tested, both exhibiting an IC50 value of 0.40 nM. ANP(103-126) and ANP(103-123) were 3 and 70 times less potent, respectively. ANP(111-126) was unable to inhibit the binding of the radioligand at a concentration of 1 microM. Several peptides unrelated to ANP were unable to inhibit the binding of the radioligand to rat olfactory bulb membranes. Membranes labeled with /sup 125/I-ANP were incubated with cross-linking agents and subjected to SDS-PAGE followed by autoradiography. A band possessing an apparent molecular mass of 116 kDa was identified. The labeling of this band was progressively decreased by increasing concentrations of unlabeled ANP(99-126) (IC50 = 0.6 nM) and by several other ANP-related peptides at nanomolar concentrations. For comparison purposes, ANP binding sites in rat aorta membranes were labeled with /sup 125/I-ANP and cross-linked using identical techniques. Three bands possessing molecular masses of 120, 72, and 62 kDa were identified. These results indicate that the ANP binding site in rat olfactory bulb membranes displays pharmacological and biochemical properties similar to peripheral ANP receptors.
- Research Organization:
- Univ. of Pennsylvania, Philadelphia
- OSTI ID:
- 6846796
- Journal Information:
- J. Neurosci.; (United States), Journal Name: J. Neurosci.; (United States) Vol. 12; ISSN JNRSD
- Country of Publication:
- United States
- Language:
- English
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59 BASIC BIOLOGICAL SCIENCES
ANIMALS
AORTA
ARTERIES
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BLOOD VESSELS
BODY
BRAIN
CARDIOVASCULAR SYSTEM
CELL CONSTITUENTS
CELL MEMBRANES
CENTRAL NERVOUS SYSTEM
CHEMICAL REACTIONS
CROSS-LINKING
DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
ELECTROPHORESIS
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPES
LABELLING
LIGANDS
MAMMALS
MEMBRANE PROTEINS
MEMBRANES
MOLECULAR WEIGHT
NERVOUS SYSTEM
NUCLEI
ODD-EVEN NUCLEI
OLFACTORY BULBS
ORGANIC COMPOUNDS
ORGANS
PEPTIDES
POLYMERIZATION
PROTEINS
RADIOISOTOPES
RATS
RECEPTORS
RODENTS
VERTEBRATES