title: Human pituitary prolactin: isolation and characterization of three isohormones with different bioassay and radioimmunoassay activities.
abstract: Three forms of human pituitary prolactin, separable at alkaline pH in a highly purified preparation, were isolated by means of column electrophoresis in agarose suspension. The most acidic component showed a significantly lower radioimmunological activity but a higher bioactivity than the other two components, which were approximately equipotent in both assays. Consequently, in both assays the most acidic component differed markedly from the other components. Amino acid analysis indicated close similarity between the three components and no size heterogeneity was observed by sodium dodecyl sulphate electrophoresis in polyacrylamide gel. A high degree of purity of the isolated components was demonstrated by analytical electrophoresis in polyacrylamide gel at alkaline pH. Runs in the same medium also revealed that the difference in electrophoretic migration velocity remained after reduction and alkylation of the isohormones. A comparison of the migrations of the S-carboxymethylated and the S-carbamidomethylated derivatives with those of the unmodified components indicated that the diversity in electrophoretical behaviour of two adjacent isohormones was consistent with a difference in one single net charge. By analytical isoelectric focusing the component of intermediate migration velocity was resolved into two distinct bands proposed to contain isohormones differing only in the exchange of one residue of aspartic acid for one of glutamic acid. The isoelectric points of the prolactin isomers were all in the range of 5.7--5.9.
mesh_terms: D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001681:Biological Assay; D010856:Columbidae; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007525:Isoelectric Focusing; D011388:Prolactin / Q000302:isolation & purification*; D011863:Radioimmunoassay
chemical_list: D000596:Amino Acids; D011388:Prolactin