An abundant and specific binding site for the novel vasodilator adrenomedullin in the rat.
Rat adrenomedullin is a novel 50-amino acid peptide with structural similarities to the calcitonin family of peptides, calcitonin, calcitonin gene-related peptide (CGRP), and islet amyloid polypeptide (IAPP). Using rat [125I]adrenomedullin, specific binding sites were demonstrated in heart, lung, spleen, liver, soleus, diaphragm, gastrocnemius, and spinal cord membranes. The highest binding was present in heart and lung, which was further characterized. These sites exhibited saturation, dissociation, and competition. In rat lung, only rat (IC50 = 5.8 nM) and human (IC50 = 94 nM) adrenomedullin competed with [125I]adrenomedullin. However, in rat heart, rat (IC50 = 0.2 nM) and human (IC50 = 4.2 nM) adrenomedullin, IAPP (IC50 = 240 nM), and CGRP (IC50 = 1050 nM) all competed with [125I] adrenomedullin. Saturation analysis revealed binding capacities and dissociation constants of 2.8 +/- 0.3 pmol/mg protein and 1.3 +/- 0.3 nM, respectively, in lung and 0.47 +/- 0.11 pmol/mg protein and 0.41 +/- 0.14 nM in heart. Comparison with [125I]CGRP- and [125I]IAPP-binding sites in lung showed that rat adrenomedullin could potently inhibit at these sites (IC50 = 5 and 6 nM, respectively). Chemical cross-linking demonstrated a major band of 83,000 mol wt in lung, diaphragm, spleen, and liver and a band of 94,000 mol wt in heart, soleus, and gastrocnemius. Thus, [125I]adrenomedullin-binding sites in rat lung are abundant and can be differentiated from binding sites in rat heart, both pharmacologically and by mol wt.
Owji
A A
AA
Department of Medicine, Royal Postgraduate Medical School, Hammersmith Hospital, London, United Kingdom.
Smith
D M
DM
Coppock
H A
HA
Morgan
D G
DG
Bhogal
R
R
Ghatei
M A
MA
Bloom
S R
SR
D053607:Adrenomedullin; D000682:Amyloid / Q000494:pharmacology; D000818:Animals; D000959:Antihypertensive Agents / Q000378:metabolism; D001665:Binding Sites; D001667:Binding, Competitive; D001921:Brain / Q000378:metabolism; D015740:Calcitonin Gene-Related Peptide / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism; D006801:Humans; D007457:Iodine Radioisotopes; D058228:Islet Amyloid Polypeptide; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D018482:Muscle, Skeletal / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism; D009928:Organ Specificity; D010455:Peptides / Q000378:metabolism*; D011869:Radioligand Assay; D051381:Rats; D058265:Receptors, Adrenomedullin; D018015:Receptors, Calcitonin Gene-Related Peptide / Q000378:metabolism*; D018000:Receptors, Peptide; D013116:Spinal Cord / Q000378:metabolism; D013154:Spleen / Q000378:metabolism; D013379:Substrate Specificity; D014665:Vasodilator Agents / Q000378:metabolism*
D000682:Amyloid; D000959:Antihypertensive Agents; D007457:Iodine Radioisotopes; D058228:Islet Amyloid Polypeptide; D010455:Peptides; D058265:Receptors, Adrenomedullin; D018015:Receptors, Calcitonin Gene-Related Peptide; D018000:Receptors, Peptide; D014665:Vasodilator Agents; D053607:Adrenomedullin; D015740:Calcitonin Gene-Related Peptide