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Preparation and Characterization of a Hormone-responsive Renal Plasma Membrane Fraction

Year: 1972
Type: article
Abstract: A renal plasma membrane fraction has been prepared by homogenization and differential centrifugation in an isotonic sucrose solution in 10 mm Tris, pH 7.5, and 1 mm ethylenediaminetetraacetate. Sedimentation in a continuous sucrose density gradient yielded a fraction that showed a striking decrease in specific activity of marker enzymes for endoplasmic reticulum and brush border, but a 2.7-fold increase in the specific activity of Na-K-ATPase. The specific activity of the mitochondrial marker, succinic cytochrome c reductase, was similar to that in the crude homogenate. The purified membrane fraction contained adenylate cyclase activity sensitive to parathyroid hormone, calcitonin, vasopressin, and isoproterenol. Only the isoproterenol-sensitive adenylate cyclase activity was inhibited by propranolol (10-5 m). Each of the three peptide-sensitive adenylate cyclase activities could be characterized by a specific anatomic distribution. The adenylate cyclase responses to the three peptide hormones showed partial additivity. The purified plasma membranes bound 125I-calcitonin (salmon), and this binding was blocked by unlabeled salmon calcitonin but not by adrenocorticotropic hormone or growth hormone. The membranes degraded 125I-salmon calcitonin to forms that were not precipitable with trichloroacetic acid and did not bind to fresh membranes.
DOI: 10.1016/s0021-9258(19)44672-3
Language: English
Open Access status: hybrid
Estimated APC paid: $3,430
OpenAlex ID: W1605412913
FWCI: 6.847
Cites: 32
Cited by: 166
Related to: 10