<I>In vitro</I> Biosynthesis of Human Chorionic Follicle Stimulating Hormone
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In order to explore the possibility that human chorionic FSH (hCFSH) may be synthesized <I>in vitro</I> by the placenta and secreted into the culture media, chorionic tissue of the first trimester was cultivated in the radioactive medium prepared by adding <SUP>3</SUP>H-proline and/or <SUP>14</SUP>C-glutamic acid. Purification of biosynthesized hCFSH from the media was carried out by a combination of Sephadex G-100 gel filtration, DEAE-cellulose chromatography and polyacrylamide disc-gel electrophoresis.<BR>Utilizing radioimmunoassay for hCFSH, incorporation of labeled amino acid into biosynthesized hCFSH was examined. <SUP>3</SUP>H-proline alone could not be incorporated easily into the biosynthesized hCFSH molecule but was predominantly incorporated into the biosynthesized hCG molecule. In a double labeling experiment with <SUP>3</SUP>H-proline and <SUP>14</SUP>C-glutamic acid, however, it was certified that hCFSH labeled with both <SUP>3</SUP>H-proline and <SUP>14</SUP>C-glutamic acid could be synthesized <I>in vitro</I>. Since hCFSH contains less proline and more glutamic acid than hCG, there may be a mutual relationship between the rate of incorporation of labeled amino acid into the hormones and the amino acid composition of the hormones.<BR>In addition, on DEAE-cellulose chromatography immunoreactive hCFSH eluted at a lower conductivity than native hCFSH and its elution profile was consistent, while immunoreactive hCG eluted at a higher conductivity than native hCG and its elution profile demonstrated a considerable variation. These results suggested that hCFSH synthesized <I>in vitro</I> might be stable, whereas hCG synthesized <I>in vitro</I> might exhibit a certain polymorphism due to charge heterogeneity.
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