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在分离过程中,TAME可稳定海胆卵的皮质和有丝分裂器。

TAME stabilizes the cortex and mitotic apparatus of the sea urchin egg during isolation.

作者信息

Kane R E

出版信息

Exp Cell Res. 1986 Feb;162(2):495-506. doi: 10.1016/0014-4827(86)90353-8.

Abstract

The synthetic substrate p-tosyl-L-arginine methyl ester (TAME) has been included in buffered EGTA media used for the isolation of the mitotic apparatus from clam eggs and also for the isolation of the cortex from sea urchin eggs. In the course of an investigation of the role of actin-fascin and actin-myosin interactions in cytokinesis, the isolation of the sea urchin egg cortex was re-examined and the stability of the cortex to lysis in a buffered EGTA medium near neutrality found to depend directly on the presence of TAME. Lysis of eggs at metaphase in this medium yielded a mixture of cortices and mitotic apparatuses (MA); MA stability under these conditions also required the presence of TAME, although a reduced pH allowed MA isolation in its absence. The action of TAME in stabilizing the actin-based structure of the cortex and the microtubule-based structure of the MA is not duplicated by other proteolysis inhibitors and this compound will also induce actin polymerization and gelation in extracts of the soluble cytoplasmic proteins of the egg under conditions where these are normally inhibited.

摘要

合成底物对甲苯磺酰-L-精氨酸甲酯(TAME)已被添加到用于从蛤卵中分离有丝分裂器以及从海胆卵中分离皮质的缓冲EGTA培养基中。在对肌动蛋白-肌动蛋白结合蛋白和肌动蛋白-肌球蛋白相互作用在胞质分裂中的作用进行研究的过程中,对海胆卵皮质的分离进行了重新检查,发现皮质在接近中性的缓冲EGTA培养基中对裂解的稳定性直接取决于TAME的存在。在此培养基中处于中期的卵裂解后产生皮质和有丝分裂器(MA)的混合物;在这些条件下MA的稳定性也需要TAME的存在,尽管在pH降低时在没有TAME的情况下也能分离出MA。TAME在稳定皮质的基于肌动蛋白的结构和MA的基于微管的结构方面的作用不能被其他蛋白水解抑制剂所替代,并且在通常会抑制这些作用的条件下,该化合物还会诱导卵的可溶性细胞质蛋白提取物中的肌动蛋白聚合和凝胶化。

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