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海胆卵中细胞内pH值与蛋白质合成速率之间的关系以及由氨引发的与pH无关的事件的存在。

The relation between intracellular pH and rate of protein synthesis in sea urchin eggs and the existence of a pH-independent event triggered by ammonia.

作者信息

Dubé F, Epel D

出版信息

Exp Cell Res. 1986 Jan;162(1):191-204. doi: 10.1016/0014-4827(86)90438-6.

Abstract

The relation between rate of protein synthesis and intracellular pH (pHi) was investigated in the eggs of the sea urchin Strongylocentrotus purpuratus. Increasing external pH (pHo) resulted in raising pHi of eggs and also in increased rate of protein synthesis. Similarly, at constant pHo, adding various concentrations of NH4Cl to eggs caused graded increases of both pHi and protein synthesis. Using various concentrations of NH4Cl at a low pHo and incubating eggs at high pHo, we compared protein synthesis under similar pHi conditions and this revealed that at least half the increased protein synthesis stimulated by NH4Cl is independent of induced rise of pHi, as also seems to be chromosome condensation which was never observed in eggs incubated at high pHoS. The additional pH-independent event triggered by NH4Cl does not appear related to elevated free Ca2+, since protein synthesis and chromosome condensation do not require external Ca2+ and no increases of free Ca2+ sufficient to activate the Ca2+-calmodulin-mediated enzyme NAD kinase occurred. Monensin disrupts intravesicular pH gradients but does not stimulate protein synthesis, indicating that this local effect, also promoted by NH4Cl, is not involved in ammonia-induced increase of protein synthesis. Using two other amines which have low pKa values, benzocaine and tricaine, we observed 2-fold increases in protein synthesis rates, even though pHi was lowered. While the exact nature of the pH-independent event(s) triggered by NH4Cl, and possibly by other amines, remains unidentified, its possible involvement in normal mitosis is stressed.

摘要

在紫海胆的卵中研究了蛋白质合成速率与细胞内pH值(pHi)之间的关系。外部pH值(pHo)的升高导致卵的pHi升高,同时蛋白质合成速率也增加。同样,在恒定的pHo下,向卵中添加不同浓度的氯化铵会导致pHi和蛋白质合成均呈梯度增加。在低pHo下使用不同浓度的氯化铵并在高pHo下孵育卵,我们比较了相似pHi条件下的蛋白质合成,结果表明,氯化铵刺激的蛋白质合成增加中至少有一半与pHi的诱导升高无关,染色体凝聚似乎也是如此,在高pHo下孵育的卵中从未观察到染色体凝聚。氯化铵引发的额外的不依赖pH的事件似乎与游离Ca2+升高无关,因为蛋白质合成和染色体凝聚不需要外部Ca2+,并且没有足以激活Ca2+ - 钙调蛋白介导的酶NAD激酶的游离Ca2+增加。莫能菌素会破坏囊泡内的pH梯度,但不会刺激蛋白质合成,这表明这种也由氯化铵促进的局部效应与氨诱导的蛋白质合成增加无关。使用另外两种pKa值较低的胺,苯佐卡因和丁卡因,我们观察到蛋白质合成速率增加了2倍,尽管pHi降低了。虽然由氯化铵以及可能由其他胺引发的不依赖pH的事件的确切性质仍未确定,但强调了其可能参与正常有丝分裂的情况。

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