Carboxymethylation of methionine residues in bovine pituitary luteinizing hormone and its subunits. Effects on the binding activity with receptor sites and interactions between subunits.
The reaction of iodoacetic acid with bovine lutropin (luteinizing hormone) at pH 3.0 was specific for methionine residues; it was slow and reached its equilibrium after 12 h at 37 degrees C. The number of modified methionine residues increased proportionately with the amount of the alkylating reagent in the reaction mixture. In the presence of a 20-fold molar excess of iodoacetic acid with respect to methionine, essentially all methionine residues in both subunits of bovine lutropin were carboxymethylated. Studies of various recombinations of modified and native alpha and beta subunits showed that methionine residues in bovine lutropin were not essential for interactions between subunits. Various recombinants were characterized by polyacrylamide-gel electrophoresis and gel filtration of Sephadex G-100. Immunological cross-reactivity by radioimmunoassay of the recombinants of modified alpha and beta subunits was relatively similar to that of the native subunits. However, the biological activity measured by receptor-site binding of the recombinants of alpha and beta chains with a total of three alkylated methionine residues was less than 5% of the activity of native lutropin. It is noteworthy that recombinants of a modified subunit and a native counterpart subunit regenerated 20-30 % of biological activity. These findings suggested that at least 1-2 methionine residues in each subunit are involved in the hormone-receptor interaction for bovine lutropin.
D000596:Amino Acids / Q000032:analysis; D001665:Binding Sites; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D007461:Iodoacetates; D007986:Luteinizing Hormone / Q000276:immunology / Q000378:metabolism*; D008715:Methionine / Q000378:metabolism*; D008745:Methylation; D011956:Receptors, Cell Surface; D013997:Time Factors
D000596:Amino Acids; D007461:Iodoacetates; D011956:Receptors, Cell Surface; D007986:Luteinizing Hormone; D008715:Methionine