Tremblais K, Oliver L, Juin P, Le Cabellec T M, Meflah K, Vallette F M
Unité INSERM 419, 9 Quai Moncousu, Nantes Cedex 01, 44035, France.
Biochem Biophys Res Commun. 1999 Jul 14;260(3):582-91. doi: 10.1006/bbrc.1999.0904.
It has been suggested that BCL-2 family members associate with certain organelles through their hydrophobic C-terminus which in the case of bcl-2, appears to play a key role in the regulation of apoptosis. We have investigated the association of bax with microsomal, nuclear and mitochondrial membranes using a cell-free system and found, contrary to bcl-2, that bax binds poorly to these organelles. Deletion of the C-terminal of bax (baxDeltaC) or exchanging the C-terminal ends of bax and bcl-XL suggests that the bax C-terminus is not an addressing/anchoring signal. In agreement with this observation, HL-60 cells transfected with either bax or baxDeltaC show no difference in sensitivity to an apoptotic signal. In the cell-free system, at low pH, bax becomes associated with mitochondria after a change of conformation, a result consistant with its structural homology with certain bacterial toxins. In HL-60 cells, as observed in the cell-free system, bax acquired a protease resistant conformation upon its translocation from the cytosol to the mitochondria after the induction of apoptosis.
有人提出,BCL-2家族成员通过其疏水的C末端与某些细胞器结合,就bcl-2而言,这似乎在细胞凋亡调控中起关键作用。我们使用无细胞系统研究了bax与微粒体、核膜和线粒体膜的结合情况,发现与bcl-2相反,bax与这些细胞器的结合能力较差。删除bax的C末端(baxDeltaC)或交换bax和bcl-XL的C末端表明,bax的C末端不是一个定位/锚定信号。与这一观察结果一致,用bax或baxDeltaC转染的HL-60细胞对凋亡信号的敏感性没有差异。在无细胞系统中,在低pH值下,bax在构象改变后与线粒体结合,这一结果与其与某些细菌毒素的结构同源性一致。在HL-60细胞中,如在无细胞系统中观察到的那样,在凋亡诱导后,bax从细胞质转移到线粒体时获得了一种抗蛋白酶的构象。