Gajko-Galicka Anna, Bielawski Krzysztof, Sredzinska Krystyna, Bielawska Anna, Gindzienski Andrzej
Department of Medical Chemistry, Medical Academy of Bialystok, Poland.
Mol Cell Biochem. 2002 Apr;233(1-2):159-64. doi: 10.1023/a:1015548131930.
The effect of the novel aromatic bisamidine 1 on protein synthesis in cell-free translational system isolated from rat livers was studied. The bisamidine 1 caused inhibition of [14C]leucine incorporation into proteins proportionally to its concentration. To establish a precise mechanism of inhibition, we evaluated the effect of the bisamidine 1 on the isolated ribosomes and purified to homogeneity elongation factors. Preincubation of the bisamidine 1 with ribosomes resulted in partial inhibition of their activity in whole elongation system. The eucaryotic elongation factor 1 (eEF-1) was not significantly affected by the bisamidine 1. In contrast to eEF-1, the bisamidine 1 preincubated with the eucaryotic elongation factor 2 (eEF-2) caused total inhibition of its activity in the translocation process. The inhibitory effect of the bisamidine 1 on eEF-2 activity was confirmed in diphtheria toxin-dependent ADP-ribosylation reaction. The results suggest a high action specificity of the bisamidine 1 as potential anticancer drug, since the primary target seems to be highly conserved protein-elongation factor 2.
研究了新型芳香双脒1对从大鼠肝脏分离的无细胞翻译系统中蛋白质合成的影响。双脒1导致[14C]亮氨酸掺入蛋白质的过程受到抑制，且与它的浓度成正比。为了确定精确的抑制机制，我们评估了双脒1对分离的核糖体和纯化至同质的延伸因子的影响。双脒1与核糖体预孵育导致其在整个延伸系统中的活性部分受到抑制。真核延伸因子1（eEF-1）未受到双脒1的显著影响。与eEF-1相反，双脒1与真核延伸因子2（eEF-2）预孵育导致其在转位过程中的活性完全受到抑制。双脒1对eEF-2活性的抑制作用在白喉毒素依赖性ADP-核糖基化反应中得到证实。结果表明双脒1作为潜在抗癌药物具有高度的作用特异性，因为其主要靶点似乎是高度保守的蛋白质延伸因子2。
