哺乳类(兔)红细胞分化因子的纯化及特性分析  

Purification and Characterization of Mammalian (Rabbit) Erythroid Differentiation Factor (EDF)

作　　者：费仁仁[1] 孙诠[2] 戴巾 朱斌 马跃 王鑫[1] 周雪峰[4] 王瑞瑜[1] 薛社普[1] 

机构地区：[1]中国医学科学院基础医学研究所,北京100005 [2]北京医科大学第三医院 [3]北京朝阳医院 [4]上海医科大学肿瘤医院

出　　处：《生物化学杂志》1992年第1期121-128,共8页

摘　　要：本文报道从哺乳动物(兔)红细胞分离、提纯红细胞分化因子(ErythroidDifferentiation Factor,简称EDF及对其性质的分析。按Bishop方法给新西兰兔注射盐酸苯肼(2.5％)0.3mL/kg体重/日连续6days致贫血,收集血液用生理盐水洗去白细胞后,得到的网织红细胞用冻融法使细胞裂解,用乙醇、氯仿选择性变性除去血红蛋白,凝胶过滤,连续经DEAE-纤维素及CM-纤维素柱层析,取得较原裂解液纯化了31 000倍的活性物质EDF,产率为9.1％,分子量15kD,在SDS-PAGE电泳图中为单一蛋白带。EDF对热相对稳定,100℃处理10min仍保持一定活性,对蛋白酶敏咸而对DNA酶I及RNA酶不敏感。EDF对体外培养细胞的生长抑制作用已在体外实验体系得到验证。EDF, derived from mammalian erythroid (reticulocytes), has been isolated and purified to apparent homogenet.New Zealand rabbits were made anemic by daily subcutaneous injection of 2.5% phenylhydrozine hydrochloride (0.3mL/kg/ day) for 6 days consecutively.Blood collected from anemic rabbits was washed by saline to remove leukocytes, and reticulocytes obtained were lysated by repeated freezing and thawing.Hemoglobin was removed by selective denaturation with ethanol and chloroform.The material was then passed over a column of Sephadex G-10O.The fraction containing the EDF was concentrated on ultrafilters and applied to a DEAE-cellulose column snd followed by chromatography on CM-cellulose, a purified protein represented a 31000 fold purification from the original lysate with an overall yield of 9.1% was achieved.The EDF was detectable as a single band in SDS-ele-ctrophoregram and with an observed molecular weight of about 15kD.EDF is relative heat stable (10 minutes at 100**********C), sensitive to protease K, but not to DNase I and RNase.The inhibitory activity of EDF on cell growth of autotransformed and tumor cells has been verified by a series of in vitro assay system.

关 键 词：网织红细胞 红细胞 分化因子 

分 类 号：R329.26[医药卫生—人体解剖和组织胚胎学]