Reaction of the histidines of prolactin with ethoxyformic anhydride. A binding site modification.
The seven histidines of bovine prolactin were modified with ethoxyformic anhydride and two classes of reactivity were apparent: 5 histidines were in the more reactive class (k = 0.097 min-1) and 2 histidines were less reactive (k = 0.011 min-1). The activity of the modified prolactins was determined by measuring their ability to bind to prolactin receptors from rabbit mammary glands. This assay showed that prolactin was fully active when 0 to 5 histidines were modified. If all 7 residues were modified, the hormone was unable to bind to its receptor. Circular dichroism studies indicated no significant differences in conformation for prolactins which had 2 to 7 histidines modified. Modification of human growth hormone and human placental lactogen with ethoxyformic anhydride resulted in a loss of the ability of these lactogenic hormones to bind to the prolactin receptor. For all three hormones, essentially full activity was recovered when the modifying group was removed by treatment with hydroxylamine. Sequence comparisons indicate that only 2 of the 3 growth hormone histidines and 2 of 7 placental lactogen histidines were homologous with histidines in bovine prolactin and that, in each case, they correspond to His-27 and His-30 in bovine prolactin. It is postulated that these residues serve to identify a portion of the binding domain of bovine prolactin.
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001681:Biological Assay; D002417:Cattle; D004047:Diethyl Pyrocarbonate; D005260:Female; D005561:Formates; D006639:Histidine; D006801:Humans; D008321:Mammary Glands, Animal / Q000378:metabolism; D010928:Placental Lactogen; D011388:Prolactin / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D011817:Rabbits; D011955:Receptors, Drug / Q000378:metabolism; D012756:Sheep; D013045:Species Specificity
D005561:Formates; D011955:Receptors, Drug; D006639:Histidine; D011388:Prolactin; D010928:Placental Lactogen; D004047:Diethyl Pyrocarbonate