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Purification and characterization of cleavage accelerating substances extracted with acid sea water from unfertilized sea urchin eggs

酸性海水未受精海胆卵裂解促进物质的纯化及表征

基本信息

批准号：
63540569
负责人：
KOJIMA Manabu
金额：
$ 1.22万
依托单位：
Toyama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1989
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63540569/
关键词：
Sea urchin eggs Cleavage accelerating substance Acid sea water extract

项目摘要

The acid sea water extract from unfertilized sea urchin eggs (egg extract:EE) can induce artificial activation, acceleration of cleavage, and animalization in sea urchin eggs. In this study, we attempted to purify and characterize cleavage accelerating factors (CAF) in EE.EE contains at least two kinds of accelerating factors; that is, a collodion bag-filterable one(MW < 12,000) and a collodion bag-non-filterable one (MW <greater than or equal>12,000). At first, we have tried to purify the former factor with low molecular weight. This collodion bag filterable factor was concentrated and fractionated by gel filtration (Bio Gel P-6 and P-2) and HPLC (ODS C-18). Thus we could obtained a very effective fraction of CAF which seemed to be a nucleotide or nuleoside (MW: <less thanOn the other hand, the collodion bag-non-filterable factor was applied to DEAE Sephadex A-50 and a fraction containing CAF was obtained. In this fraction, a small amount of proteins was detected, so that the factor seemed to be a nucleotide or nucleoside, binding some proteins.Furthermore, it was found that the collodion bag-non- filterable factor contains a cleavage inhibiting factor which stops cytokinesis completely. Therefore, it seems that a cell cycle is regulated by balance of two antipodal actions of such factors, although this inhibiting factor should be studied more precisely.It was also revealed that neither cytoplasmic free Ca^+ concentration nor cytoplasmic pH are not raised by EE-treatment, but the content of arginine phosphate is increased to the level in fertilized eggs. Such a changes may have some relationship to acceleration of cleavage induced by EE-treatment.

未受精的海胆卵的酸性海水提取物（卵提取物：EE）可以诱导海胆卵的人工激活，加速卵裂和动物化。EE中至少含有两种加速因子，即胶棉袋过滤型（MW <12，000）和胶棉袋非过滤型（MW <greater than or equal>12，000）。首先，我们尝试用低分子量的方法纯化前者。通过凝胶过滤（Bio Gel P-6和P-2）和HPLC（ODS C-18）浓缩和分级该火棉胶袋可过滤因子。另一方面，将胶棉袋不可滤因子应用于DEAESephadexA-50，得到了含有CAF的组分。在该组分中，检测到少量的蛋白质，因此该因子似乎是核苷酸或核苷，与某些蛋白质结合。此外，发现火棉胶袋不可过滤因子含有完全停止胞质分裂的切割抑制因子。因此，细胞周期似乎是由这两种因子的反足作用的平衡来调节的，尽管这种抑制因子还需要更精确地研究。还表明，EE处理既不能提高细胞质游离Ca^+浓度，也不能提高细胞质pH值，但精氨酸磷酸盐的含量增加到受精卵的水平。这种变化可能与EE处理诱导的卵裂加速有关。