Bovine Cerebellum Endothelin Receptor: Solubilization and Indentification
- 1 June 1990
- journal article
- research article
- Published by The Endocrine Society in Endocrinology
- Vol. 126 (6) , 3218-3222
- https://doi.org/10.1210/endo-126-6-3218
Abstract
Endothelin receptors were solubilized from bovine cerebellum membrane preparations in an active form by using the zwitterionic detergent CHAPS, [3-(3-cholamidoropyl)dimehtylammonio]-1-propane sulfonic acid. The solubilized receptors displayed high affinity, saturability, and specificity. The dissociation constant (Kd) for endothelin was 7 .+-. 2 nM, and the number of binding sites was 600 .+-. 200 fmol/mg protein. These results are similar to those obtained for the membrane-bound receptor and suggest that during solubilization the binding characteristics of the receptor are preserved. Attempts to purify the solubilized receptors in an active form using affinity chromatography techniques, i.e. Affi-gel 15 column coupled to endothelin, were not successful. Nevertheless, identification of the receptors was achieved by affinity chromatography of the solubilized proteins and subsequent iodination. Autoradiographic analysis of sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a major protein with an apparent mol wt of 50 kD. Taken together with our previous findings, this result suggests that the 50-kD band represents the endothelin receptor.This publication has 15 references indexed in Scilit:
- Endothelin 1, an endothelium-derived peptide, is expressed in neurons of the human spinal cord and dorsal root ganglia.Proceedings of the National Academy of Sciences, 1989
- Two distinct types of endothelin receptors are present on chick cardiac membranesBiochemical and Biophysical Research Communications, 1989
- Primary structure, synthesis, and biological activity of rat endothelin, an endothelium-derived vasoconstrictor peptide.Proceedings of the National Academy of Sciences, 1988
- Cellular mechanism of action by a novel vasoconstrictor endothelin in cultured rat vascular smooth muscle cellsBiochemical and Biophysical Research Communications, 1988
- Solubilization and purification of rat pituitary gonadotropin-releasing hormone receptor.Journal of Biological Chemistry, 1986
- Purification of insulin receptor with full binding activity.Journal of Biological Chemistry, 1983
- Solubilization of active prolactin receptors by a nondenaturing zwitterionic detergent.Journal of Biological Chemistry, 1982
- The beta-adrenergic receptor: rapid purification and covalent labeling by photoaffinity crosslinking.Proceedings of the National Academy of Sciences, 1982
- Purification and properties of the human placental insulin receptor.Journal of Biological Chemistry, 1981
- Solubilization of active opiate receptors.Proceedings of the National Academy of Sciences, 1980