Maruo T, Cohen H, Segal S J, Koide S S
Proc Natl Acad Sci U S A. 1979 Dec;76(12):6622-6. doi: 10.1073/pnas.76.12.6622.
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 beta 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 beta subunit and hCG beta subunit COOH-terminal peptide were also found in the extracts. No free alpha subunit was detected. The CG-like 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.
从一种暂命名为隐球菌始祖菌培养物的丙酮粉制剂中提取的物质,经用人绒毛膜促性腺激素(hCG)β亚基羧基末端肽抗血清进行放射免疫测定以及用牛黄体膜进行放射受体测定,发现含有绒毛膜促性腺激素(CG)样因子。提取物中蛋白酶的可能干扰已被排除。提取物中还发现了与hCGβ亚基和hCGβ亚基羧基末端肽抗血清发生反应的免疫反应性物质。未检测到游离的α亚基。通过在葡聚糖凝胶G - 100、伴刀豆球蛋白A - 琼脂糖和二乙氨基乙基葡聚糖A - 50上进行色谱分离,对CG样因子进行了纯化。该因子被伴刀豆球蛋白A - 琼脂糖吸附,表明它含有甘露糖和葡萄糖部分。在葡聚糖凝胶G - 100上,该因子在与标准hCG相同的位置洗脱。当进行十二烷基硫酸钠/聚丙烯酰胺凝胶电泳时,它解离成两条带;这两条带分别对应于各自的标准hCG亚基。通过大鼠子宫重量测定法和卵巢重量测定法测定,纯化因子的生物活性分别相当于380(95%置信限:320 - 490)和880(780 - 1020)国际单位/毫克。从目前的结果可以得出结论,一种微生物产生了一种具有生物活性且理化性质与hCG相似的糖蛋白。