Solubilization and molecular weight estimation of atrial natriuretic factor receptor from bovine adrenal cortex.
Receptors for atrial natriuretic factor (ANF) have been solubilized with 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate from bovine adrenal cortex and characterized. The detergent extract retained specific high-affinity binding sites for 125I-ANF. Scatchard analysis of the equilibrium binding data revealed a single class of binding site with a K-d of 1.8 nM and a maximum binding capacity of 2.5 pmol/mg of protein. The size of the 125I-ANF X receptor complexes was estimated to be 140,000 daltons by gel filtration on TSK gel G3000SW. Affinity labeling followed by electrophoresis under nonreducing conditions and autoradiography also revealed a single band of a similar size (Mr = 130,000); this band, however, migrated as a Mr = 70,000 species under reducing electrophoretic conditions. These results indicate that the ANF receptor, having a Mr of 130,000 - 140,000, is composed of disulfide-linked subunits and the ANF-binding site is located on the 70-kDa component.
Hirose
S
S
Akiyama
F
F
Shinjo
M
M
Ohno
H
H
Murakami
K
K
D000302:Adrenal Cortex / Q000032:analysis*; D000345:Affinity Labels / Q000378:metabolism; D000818:Animals; D009320:Atrial Natriuretic Factor; D002417:Cattle; D002793:Cholic Acids; D002851:Chromatography, High Pressure Liquid; D008970:Molecular Weight; D009124:Muscle Proteins / Q000378:metabolism; D017461:Receptors, Atrial Natriuretic Factor; D011956:Receptors, Cell Surface / Q000032:analysis*; D012995:Solubility
D000345:Affinity Labels; D002793:Cholic Acids; D009124:Muscle Proteins; D011956:Receptors, Cell Surface; D009320:Atrial Natriuretic Factor; D017461:Receptors, Atrial Natriuretic Factor; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate