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Solubilization of receptor for pituitary adenylate cyclase activating polypeptide from bovine brain
1Tsuruba Research Laboratories, Takeda Chemical Industries, Ltd., Ibaraki, Japan.
Biochemical and Biophysical Research Communications
|October 30, 1990
Abstract:
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.