通过二乙氨基乙基纤维素色谱法纯化海胆精子结合蛋白。

Purification of sea urchin sperm bindin by DEAE-cellulose chromatography.

作者信息

Vacquier V D

出版信息

Anal Biochem. 1983 Mar;129(2):497-501. doi: 10.1016/0003-2697(83)90583-3.

Abstract

A procedure for purifying bindin from sperm of the sea urchin Strongylocentrotus purpuratus is presented in detail. The impure bindin, dissolved in 4 M urea, 50 mM sodium phosphate, pH 6.6, is adsorbed to DEAE-cellulose and eluted wit 4 M urea, 650 mM sodium phosphate, pH 6.6. The purified bindin is not contaminated with tubulin or histone HI. A precipitate of this DEAE-purified bindin, made by dialysis into Ca2+-free seawater and natural seawater, is a species-specific agglutinin of unfertilized eggs. This method of obtaining consistently pure preparations of bindin will aid in the analysis of its role in fertilization.

摘要

详细介绍了一种从紫海胆精子中纯化结合素的方法。将溶解于4M尿素、50mM磷酸钠（pH 6.6）中的粗制结合素吸附到DEAE - 纤维素上，并用4M尿素、650mM磷酸钠（pH 6.6）洗脱。纯化后的结合素未被微管蛋白或组蛋白H1污染。通过透析到无Ca2 +的海水和天然海水中制成的这种DEAE纯化结合素沉淀，是未受精卵的种特异性凝集素。这种始终能获得纯结合素制剂的方法将有助于分析其在受精过程中的作用。