Solubilization of receptor for pituitary adenylate cyclase activating polypeptide from bovine brain.
A receptor for pituitary adenylate cyclase activating polypeptide (PACAP) was solubilized from bovine brain membranes with 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate. Saturable binding experiments using [125I]PACAP27 revealed that the solubilized extract contained a single class of binding sites with a Kd of 200 pM and a Bmax of 0.9 pmol/mg protein. The Kd did not change significantly after the solubilization. Competitive binding experiments confirmed that the solubilized receptor retained a specificity for PACAP. Thus, the solubilization of the receptor in an active form was successfully achieved by the present procedure.
Masuda
Y
Y
Tsuruba Research Laboratories, Takeda Chemical Industries, Ltd., Ibaraki, Japan.
Ohtaki
T
T
Kitada
C
C
Tsuda
M
M
Arimura
A
A
Fujino
M
M
D000818:Animals; D001667:Binding, Competitive; D001921:Brain / Q000378:metabolism*; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D002793:Cholic Acids; D003902:Detergents; D007700:Kinetics; D011956:Receptors, Cell Surface / Q000302:isolation & purification* / Q000378:metabolism; D051236:Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide; D011979:Receptors, Pituitary Hormone; D012995:Solubility
D002793:Cholic Acids; D003902:Detergents; D011956:Receptors, Cell Surface; D051236:Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide; D011979:Receptors, Pituitary Hormone; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate