摘要
从螺旋藻体中通过ＤＥＡＥ纤维素层析、硫酸铵分部沉淀和葡聚糖Ｇ７５凝胶过滤得到一种具有离体情况下刺激红系细胞集落生成的功能蛋白质．ＳＤＳ—ＰＡＧＥ测分子量为１５０００道尔顿，等电点为４．８，含有藻蓝胆素．该蛋白命名为螺旋藻１５０００蛋白（Ｓｐｉｒｕｌｉａｐｌａｔｅｎｓｉｓ１５０００，ＳＰ—１５０００）．
The differentiation and maturation of erythrocytes. are regulated by severalfactors in vivo, among which erythropoietin (EPO) is known as the most improtantone.The activity of EPO could be determined by counting CFU-E in vitro.In this article, a phydocyanin (PS-15000) isolated from a species of blue-green alge(Spirlina Platensis) with high erythropoietinlike activity was reported. The spirulia platestscells suspended in 20 Volum of 50mM pH 7.3 PBS containing 0.1 % Triton X-100; freezing and thawing 3 tAnes for the cell broken, after centrifugalized at 10,000 rpm for 30min, the supernants were separated on DE-52 cellulose chromatography.The fraction of10 mM pH6.0 PBS containing 0.3N NaCI was fractionated with amminium sulfat in 80%saturation, dissolved the precipitate in water and dialyzed against 10mM .pH 7. 3 PBSovernight.The dialyzed sample was applied on a sephadex G75 column.The collected main.fraction of the gel filtration shows at single band with molecular weight 15,000 Dolton onSDS- PAGE, and PI 4. 8 on gel isoelectric focusing electrophresis, containingphycocyanobilin.EPO-like activity of SP-15,000 was determined with liver cellsof 13 days fetal mice .asthe target cells, plated the GCM-cellulose soft gel as culture mediun in 35mm Petri deshesand incubated at 37 °C, 5% CO2for 72 hours.Then we counted .the CEU-E with lightmicroscope , ane a colony containing at least 8. cells was defined as a CFU-E, Thestand ard.of EPO was provided by English National Institute for biological standrards and con -trol The specific activity fo SP-15,000 was higher than 70: 000u / mg protien.This proteinfrom alge which regulates the proliferation and differentiation of erythrocytes will be very promising .
出处
《南京大学学报（自然科学版）》
CSCD
1994年第2期377-380,共4页
Journal of Nanjing University（Natural Science）
关键词
螺旋藻
蛋白质
红系细胞集落
Spirulina platensis
Erythropoietin (EPO)
Erythroid colony forming unit (CFU-E) .
