Boll Werner, Ehrlich Marcelo, Collier R John, Kirchhausen Tomas
Department of Cell Biology, The CBR Institute for Biomedical Research, Harvard Medical School, Boston, MA 02115, USA.
Eur J Cell Biol. 2004 Jul;83(6):281-8. doi: 10.1078/0171-9335-00373.
Internalization and traffic to acidic endosomes of anthrax lethal factor (LF) and protective antigen (PA), bound to the anthrax toxin receptor (ATR), is required for LF translocation into the cytosol, where it can elicit its toxic effects. Dynamin is required for clathrin-mediated endocytosis, and long-term disruption of dynamin function blocks internalization of PA. We have used LFn-DTA, a surrogate of LF consisting of the N-terminal domain of LF fused to the catalytic subunit of diphtheria toxin, to differentiate the effects of acute and long-term block of dynamin function on LFn-DTA toxicity. Both forms of interference reduce LFn-DTA toxicity only partially, consistent with alternative routes for LFn-DTA endocytosis. In contrast, a long-term block of dynamin activity results in a further interference with LFn-DTA toxicity that is consistent with an altered endosomal environment, probably an increase in endosomal pH.
炭疽致死因子(LF)和保护性抗原(PA)与炭疽毒素受体(ATR)结合后,其内化并转运至酸性内体是LF转运至胞质溶胶所必需的,在胞质溶胶中LF可发挥其毒性作用。发动蛋白是网格蛋白介导的内吞作用所必需的,长期破坏发动蛋白功能会阻断PA的内化。我们使用了LFn-DTA(一种由LF的N端结构域与白喉毒素催化亚基融合而成的LF替代物)来区分发动蛋白功能急性和长期阻断对LFn-DTA毒性的影响。两种干扰形式都只是部分降低了LFn-DTA的毒性,这与LFn-DTA内吞作用的替代途径一致。相比之下,长期阻断发动蛋白活性会进一步干扰LFn-DTA的毒性,这与内体环境改变一致,可能是内体pH值升高。