大鼠肝细胞溶质中抗有丝分裂物质——肝细胞溶质coronin的纯化及某些特性

Purification and some characteristics of liver cytosol cornin, an antimitotic substance from rat liver cytosol.

作者信息

Inaba K, Doi A, Nisida I

出版信息

Acta Med Okayama. 1977 Jun;31(3):203-9.

Abstract

Further purification and characterization are reported on rat cytosol cornin (RLCC), an antimitotic substance. Fraction I (purified RLCC) was purified more than 10-fold from crude RLCC with Sephadex G-50 column chromatography and showed a remarkable inhibitory effect on division of inseminated sea urchin eggs and mouse fibroblast cells. Fraction I was observed as one spot, and the molecular weight was estimated to be about 25,000 by thin layer gel filtration. Fraction I contained protein (92%) and RNA (8%), but the antimitotic activity was scarcely affected by treatment by pancreatic RNase. The protein of Fraction I was separated into two bands by SDS-polyacrylamide gel electrophoresis, and the molecular weight was estimated as 10,000 and 15,000, respectively. The 50% inhibition dose of Fraction I on the first division of inseminated sea urchin eggs and on proliferation of mouse L cells was about 2.5 X 10(-5) g/ml and 5 X 10(-4) g/ml, respectively. The yield of fraction I was about 35 mg from 100 g rat liver.

摘要

本文报道了对一种抗有丝分裂物质——大鼠细胞质coronin（RLCC）的进一步纯化和特性研究。通过Sephadex G - 50柱层析从粗RLCC中获得了纯化超过10倍的组分I（纯化的RLCC），其对受精海胆卵和小鼠成纤维细胞的分裂显示出显著的抑制作用。组分I在薄层层析中呈现为一个斑点，通过薄层层析凝胶过滤法估计其分子量约为25,000。组分I含有蛋白质（92%）和RNA（8%），但其抗有丝分裂活性几乎不受胰核糖核酸酶处理的影响。通过SDS - 聚丙烯酰胺凝胶电泳将组分I的蛋白质分离为两条带，其分子量分别估计为10,000和15,000。组分I对受精海胆卵第一次分裂和小鼠L细胞增殖的50%抑制剂量分别约为2.5×10⁻⁵ g/ml和5×10⁻⁴ g/ml。从100 g大鼠肝脏中获得的组分I产量约为35 mg。