Photoaffinity labeling of the canine renal receptor for parathyroid hormone.
Studies were undertaken to identify and characterize components of the parathyroid hormone receptor. An analog of the bovine parathyroid hormone(1-34) sequence was derivatized at the 23-tryptophan position with the photoreactive reagent 2-nitro-5-azidophenylsulfenyl chloride. 2-Nitro-5-azidophenylsulfenyl-bovine parathyroid hormone (NAPS-bPTH) analog retained full biological activity with respect to receptor binding and activation of adenylate cyclase in canine renal cortical plasma membranes. 125I-bPTH(1-34) analog was also derivatized without loss of receptor-binding activity. When 125I-NAPS-bPTH(1-34) analog was incubated with canine renal plasma membranes and then photolyzed, at least two 125I-labeled membrane components were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The incorporation of 125I radioactivity into one of these components (Mr congruent to 60,000) was inhibited when incubation and photolysis were performed in the presence of excess, unlabeled bPTH(1-34). Furthermore, photolysis of membranes in the presence of NAPS-bPTH(1-34) analog led to activation of adenylate cyclase which persisted following washing to remove noncovalently bound peptide, suggesting a functional, covalent hormone-receptor complex had been formed. We conclude that the M r congruent to 60,000 membrane component may be a part of the renal receptor for parathyroid hormone.
D000262:Adenylyl Cyclases / Q000378:metabolism; D000345:Affinity Labels / Q000138:chemical synthesis; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001386:Azides; D002462:Cell Membrane / Q000378:metabolism; D004285:Dogs; D004789:Enzyme Activation; D007668:Kidney / Q000378:metabolism*; D007672:Kidney Cortex / Q000378:metabolism; D010281:Parathyroid Hormone / Q000138:chemical synthesis / Q000378:metabolism* / Q000494:pharmacology*; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000378:metabolism*; D018016:Receptors, Parathyroid Hormone
D000345:Affinity Labels; D000596:Amino Acids; D001386:Azides; D010281:Parathyroid Hormone; D011956:Receptors, Cell Surface; D018016:Receptors, Parathyroid Hormone; C033885:parathyroid hormone, 2-nitro-5-azidophenylsulfenyl-, bovine; D000262:Adenylyl Cyclases