Production of choriogonadotropin-like factor by a microorganism.

Abstract

Extracts from an acetone powder preparation of a culture of a microorganism tentatively named Progenitor cryptocides contain choriogonadotropin (CG-like factor as determined by radioimmunoassay with antiserum to human (h)CG # subunit COOH-terminal peptide and radioreceptor assay with bovine corpus luteum membranes. Possible interference by proteases in the extracts was excluded. Immunoreactive materials reacting with antisera to hCG 0 subunit and hCG P subunit COOH-terminal peptide were also found in the extracts. No free a subunit was detected. The CClike factor was purified by chromatography on Sephadex G-100, concanavalin A-Sepharose, and DEAE-Sephadex A-50. The factor was adsorbed by concanavalin A-Sepharose, suggesting that it contains mannose and glucose moieties. The factor was eluted at the same position as standard hCG on Sephadex G-100. It dissociated into two bands when subjected to sodium dodecyl sulfate/polyacrylamide gel electrophoresis; the bands corresponded to the respective standard hCG subunits. The biological activity of the purified factor as determined by the rat uterine weight assay and the ovarian weight assay was equivalent to 380 (95% confidence limits: 320-490) and 880 (780-1020) international units/mg, respectively. It can be concluded from the present results that a microorganism produces a glycoprotein that is biologically active and has physicochemical properties similar to those of hCG.Human (h-) choriogonadotropin (CG) is a glycoprotein hormone normally produced by the placenta and considered to be a specific hormone of pregnancy and trophoblastic neoplasms. However, it has been reported that nontrophoblastic malignant tumors (1, 2) and normal human tissues (3) produce CG Related to the problem of extraplacental production of CG, there are reports suggesting that CG is produced by a microorganism (5, 6). However, in the measurements in these studies, possible interference by proteases in the bacterial extracts was not excluded. In the present study, we will show that a CG-like factor is produced by a microorganism and that the observed results are not due to proteases. The factor has been purified and found to be biologically active, and it has immunological and physicochemical properties similar to those of hCG. MATERIALS AND METHODS Bacterial Extracts. A microorganism tentatively namedProgenitor cryptocides was obtained from V. W.-C. Livingston (5). Cultures of the microorganism were maintained on Muller-Hinton slants. Inoculation of 250 ml of trypticase/yeast extract medium in a 500 ml Erlenmeyer-flask was performed with a single colony from the Muller-Hinton slant. To a 28-liter fermenter, 20 liters of trypticase/yeast extract medium was added and the fermenter was inoculated with an overnight growth of the organism in the 250 ml of medium described above. After incubation for 120 hr at 37°C, the entire culture content was harvested and an acetone-dried powder was prepared according to a prescribed method (5), by J. Majnarich, Biomedical Research Laboratory. Twenty ...

Corticotropin and β-endorphin-like materials are native to unicellular organisms

“…Almost all ofthe immunoreactive ,3endorphin detected exhibited a molecular weight >28,000. The distribution of immunoreactive (3-endorphin was qualitatively similar to that of the multiple high molecular weight forms seen in the column Migration, cm DISCUSSION Recent studies have demonstrated material similar to mammalian insulin in unicellular eukaryotes and prokaryotes (a protozoan, two fungi, and a bacterium) (3), as well as material similar to the placental form of human chorionic gonadotropin in a bacterium (15). In the present study, we found material in extracts ofT.…”

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Effects of antibodies to choriogonadotropin in malignant growth

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There is no turning back on the paradigm shift on the actions of human chorionic gonadotropin and luteinizing hormone

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