Ng T B, Lo L L, Chan W Y
Department of Biochemistry, Faculty of Medicine, Chinese University of Hong Kong, Shatin, N.T.
Biochem Int. 1992 Dec;28(6):999-1007.
The pituitaries of mice, hamsters, guinea pigs and rats were extracted with Tris-Cl buffer and the extracts were chromatographed on Concanavalin A (ConA)-Sepharose into unadsorbed ConA I and adsorbed ConA II fractions. The ConA I fraction was subjected to gel filtration on Sephadex G-100 and fractionated into an unretarded peak and several retarded peaks. The peak with a molecular weight of approximately 40,000 (designated ConAI Sephadex fraction II) was then subjected to ion exchange chromatography on CM-cellulose and fractionated into an unadsorbed CM I and an adsorbed CM II fraction. The ConA II fraction was fractionated by ion exchange chromatography on CM-cellulose into CM I and CM II fractions. The ConA II CM II fraction was the chromatographic fraction which exhibited the highest potency in stimulating testosterone production by isolated rat Leydig cells. Its activity was much higher than the corresponding ConA I Sephadex fraction II CM II fraction which differed chromatographically only by non-adsorption on ConA-Sepharose. The ConA II CM II fraction manifested cross reactivity in a rat luteinizing hormone (LH) radioimmunoassay. The guinea pig pituitary ConA II CM II fraction also cross-reacted in a rat thyroid stimulating hormone radioimmunoassay. The ConA II fractions of hamster and guinea pig pituitary extracts demonstrated follicle stimulating hormone (FSH) activity while the corresponding ConA I fractions did not. The results suggest that the ConA II/ConA II CM II fraction contained most of the FSH and LH activities present in the pituitary extract.
用Tris-Cl缓冲液提取小鼠、仓鼠、豚鼠和大鼠的垂体,提取物在伴刀豆球蛋白A(ConA)-琼脂糖上进行色谱分离,得到未吸附的ConA I和吸附的ConA II组分。ConA I组分在Sephadex G-100上进行凝胶过滤,分离为一个无滞留峰和几个滞留峰。然后将分子量约为40,000的峰(称为ConAI Sephadex组分II)在CM-纤维素上进行离子交换色谱分离,分为未吸附的CM I和吸附的CM II组分。ConA II组分通过在CM-纤维素上进行离子交换色谱分离为CM I和CM II组分。ConA II CM II组分是在刺激分离的大鼠睾丸间质细胞产生睾酮方面表现出最高效力的色谱组分。其活性远高于相应的ConA I Sephadex组分II CM II组分,这两种组分在色谱上的差异仅在于在ConA-琼脂糖上的非吸附性。ConA II CM II组分在大鼠促黄体生成素(LH)放射免疫测定中表现出交叉反应性。豚鼠垂体ConA II CM II组分在大鼠促甲状腺激素放射免疫测定中也有交叉反应。仓鼠和豚鼠垂体提取物的ConA II组分表现出促卵泡激素(FSH)活性,而相应的ConA I组分则没有。结果表明,ConA II/ConA II CM II组分包含垂体提取物中存在的大部分FSH和LH活性。