- 作者: 陳正繹; 張文章; 趙壯飛
- 作者服務機構: 國防醫學院生物及解剖學科; 國立臺灣大學理學院生亡科學研究所及中央研究院生物化學研究所
- 中文摘要:
17-β-經基類固醇去氫?是種廣泛分佈的組織性?,主要功能是催化雌性素中雌素酮與雌二醇間的轉換。胎盤中
含有大量的此類酵素,人類胎盤經過一系列磨碎離心、硫胺沈殿、及68°C加熱處理步驟後,再經過親和性層析,反向高
效率層析或製備性膠體電泳,可得到純化之蛋白質,在SDS聚丙烯胺膠體電泳上,呈現分子量約34 kDa的單一色帶。經
過非變性不連續膠體電泳、酵素染色法及高效率分子篩層析,以及二次元雙向電泳測定其具活性狀況下為分子量68
kDa之雙元體,並具有三種不同電性之異構物,大量製備之純化蛋白質經免疫紐西蘭大白免得到抗血清,由酵素吸附免
疫分析,螢光免疫組織化學法,西方墨漬法及免疫沉搬法,確定誘導得之抗血清確實對人類胎盤17-β-羥基類固醇去
氫?具專一性。
務 - 英文摘要: 17-β-Hydroxysteroid dehydrogenase was purified from human placenta. The purification processincluded the following steps: 50% saturated ammonium sulfate precipitation; heat treatment; affinity columnchromatography; and preparative SDS gel electrophoretic elution. This enzyme was also characterized byHPLC gel filtration and two dimensional gel electrophoresis with isoelectrofocusing. The results indicatethat the molecular weight of these enzymes in their native condition is around 68 kDa and 34 kDa in SDSPAGE. Therefore, the active form of the enzyme is an identical dimmer in nature. There are three chargedisomers as demonstrated by isoelectrofocusing. The antiserum against the 17-β-HSD was induced by injection of purified human placenta 17-β-HSDin rabbits. The antiserum was collected and characterized by ELISA, immunohistochemistry, immunoblotand immunoprecipitation tests. The results showed that the antibody titer was over 1:12,800, and specificityagainst human placenta 17-β-HSD was also observed.
- 中文關鍵字: 17-β-HSD; antiserum; human lacenta
- 英文關鍵字: --