细菌(Pseudomonas maltophilia)之绒毛膜促性腺激素(hCG)结合物质之研究

 细菌(Pseudomonas moltophilia)与hCG及LH有特异的亲和力,实验发现,细菌之生长曲线与hCG结合活性成平行关系,96小时达高峰,细菌之培养液中含有可溶性结合蛋白,该蛋白经硫酸铵沉淀(80％饱和度)、Sephadex G-100柱层析、DEAE-纤维素柱0.5mol/L NaCl梯度洗脱,再过Sepharose CL-AB柱,收集之活性部分经SDS电泳测得其分子量为70,000,凝胶层析测Stokes radius为41A,Schiff氏染色未见着色带。

A STUDY ON THE SPECIFIC HUMAN CHORIONIC GONADOTROPIN (HCG) BINDING TO PSEUDOMONAS MALTOPHILIA

Human Chorionic Gonadotropin (hCC) receptor is one of the eucayotic cell membrane proteins. Subsequent outgrowth of the isolated strains in liquid media demonstrated that the bacteria P maltophilia was capable of binding I25I-hCG with high affinity. The hCG binding substances from P .maltophilia required 96 hrs to reach equilibrium at 30℃ in trypiticas Soy broth culture. Experiments demonstrated the culture media contained solubilized form of the bacteria hCG binding protein. Effort has been made to isolate this hCG binding substances.The ammonium sulfate precipitates of the bacteria culture medium was chromatographed on Sephadex G-100 column. The binding activity was present in the peak A, B.Gel filtraton of peak A, B from Sephadex G-100 passed through a DEAE-Cellulose column. The activity fraction from DEAE-Cellulose was further purified on a Sepharose CL-4B column. The elution position from the peak A of the major portion of the binding activity corresponded to a protein of 41 A stokes radius and appeared as a band in SDS gel electrophoresis with approximate M.W of 70,000. When stained with Schiff's reagent the gel showed negative result.