The role of the methionine residues in the structure and function of parathyroid hormone.
Forms of the biologically active N-terminal fragment of bovine parathyroid hormone oxidized at methionine 8, methionine 18, and both positions were prepared, separated from one another, and characterized as described earlier for the native hormone (A. L. Frelinger and J. E. Zull, (1984) J. Biol. Chem. 259, 5507). The biological properties of the oxidized forms were compared to those of the native hormone, using the renal membrane adenylyl cyclase assay. Oxidation at position 18 produced full agonists of the hormone with slightly reduced potency. Oxidation at position 8 produced partial agonists of greatly reduced potency. Oxidation at both positions produced partial agonists of even lower potency. Thus, methionine 8 is implicated both in binding and in activation of adenylyl cyclase, but methionine 18 is implicated only in binding. Further study showed that oxidation of both residues is dependent on the pH, ionic strength, and polarity of the solvent. However, methionine 8 is less easily oxidized than methionine 18. This difference is eliminated in 3 M guanidine-HCl with 1-34 and in 6 M guanidine-HCl with 1-84. On the other hand the difference in reactivity is greatly increased in high ionic strength, with methionine 8 becoming much less reactive. These results suggest that the methionine residues are important in the biologically active conformation of parathyroid hormone and that methionine 8 is less accessible than methionine 18 under certain conditions. These conclusions are discussed in the context of a specific model for the folding of parathyroid hormone.
D000262:Adenylyl Cyclases / Q000378:metabolism; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D002417:Cattle; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D000432:Methanol / Q000494:pharmacology; D008715:Methionine / Q000502:physiology*; D010084:Oxidation-Reduction; D010281:Parathyroid Hormone / Q000378:metabolism / Q000502:physiology*; D011487:Protein Conformation
D000596:Amino Acids; D010281:Parathyroid Hormone; D008715:Methionine; D000262:Adenylyl Cyclases; D000432:Methanol