Solubilization and molecular characterization of the atrial natriuretic peptide (ANP) receptor in human platelets: Comparison with ANP receptors in rat tissues

Journal Article · Fri Feb 01 04:00:00 EST 1991 · Journal of Clinical Endocrinology and Metabolism; (USA)

DOI:https://doi.org/10.1210/jcem-72-2-484· OSTI ID:6062311

Schiffrin, E L; Carrier, F; Thibault, G; Deslongchamps, M ^[1]

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1. Clinical Research Institute of Montreal, Quebec (Canada)

We have previously demonstrated the presence of binding sites for atrial natriuretic peptide (ANP) in human platelets. These sites have pharmacological characteristics similar to those of rat vascular smooth muscle. They are subject to regulation by circulating levels of ANP in plasma, varying inversely with the latter after high sodium intake, in arterial hypertension and congestive heart failure. We have now solubilized these platelet receptors with the nonionic detergent Triton X-100 (0.6%). The preparations were incubated with (125I)ANP in the presence of increasing concentrations of ANP-(99-126), ANP-(101-126), ANP-(103-126), and ANP-(103-123). The order of potency of these peptides to displace (125I)ANP was similar for the solubilized and particulate receptor. Bound (125I)ANP was covalently cross-linked to the receptor with 5 mM disuccinimidyl suberate. Autoradiography of the sodium dodecyl sulfate-polyacrylamide gel showed that (125I)ANP specifically interacts with a 125-kDa membrane component, some of which may be reduced by 2% mercaptoethanol or 10 mmol/L dithiothreitol to a 70-kDa species. A small proportion of a 70-kDa peptide is also found under nonreducing conditions. The concentration of ANP-(99-126) that inhibits binding of (125I)ANP by 50% to both the 125-kDa and the 70-kDa species was 0.1 nM, while that for ANP-(103-123) was 3 nM. The internally ring-deleted analog Des(Gln116,Ser117,Gly118,Leu119,Gly120)ANP -(102-121) or C-ANP displaced with equal potency ANP binding to the high and low mol wt (Mr) bands, as also found in cultured rat vascular smooth muscle cells, but not in the mesemteric arteries these cells are derived from. In the latter, C-ANP displaced only binding from the lower Mr band. These results show that the ANP receptor in human platelets is heterogeneous.

• OSTI ID: 6062311
• Journal Information: Journal of Clinical Endocrinology and Metabolism; (USA), Journal Name: Journal of Clinical Endocrinology and Metabolism; (USA) Vol. 72:2; ISSN JCEMA; ISSN 0021-972X
• Country of Publication: United States
• Language: English

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ANIMALS
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BLOOD PLATELETS
BODY FLUIDS
CELL CONSTITUENTS
CELL MEMBRANES
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DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
INTERMEDIATE MASS NUCLEI
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ISOTOPES
KINETICS
MAMMALS
MAN
MATERIALS
MEMBRANE PROTEINS
MEMBRANES
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
PEPTIDES
PRIMATES
PROTEINS
RADIOISOTOPES
RATS
REACTION KINETICS
RECEPTORS
RODENTS
SOLUBILITY
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