Fabre Lucien, Santelli Eugenio, Mountassif Driss, Donoghue Annemarie, Biswas Aviroop, Blunck Rikard, Hanein Dorit, Volkmann Niels, Liddington Robert, Rouiller Isabelle
Department of Anatomy and Cell Biology, McGill University, Montréal, Québec H3A 0C7, Canada Groupe de Recherche Axé sur la Structure des Protéines (GRASP), Groupe d'Étude des Protéines Membranaires (GÉPROM), McGill University, Montréal, Québec H3A 0C7, Canada.
Bioinformatics and Structural Biology Program, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA 92037.
J Gen Physiol. 2016 Oct;148(4):313-24. doi: 10.1085/jgp.201611617.
Anthrax toxin comprises three soluble proteins: protective antigen (PA), lethal factor (LF), and edema factor (EF). PA must be cleaved by host proteases before it oligomerizes and forms a prepore, to which LF and EF bind. After endocytosis of this tripartite complex, the prepore transforms into a narrow transmembrane pore that delivers unfolded LF and EF into the host cytosol. Here, we find that translocation of multiple 90-kD LF molecules is rapid and efficient. To probe the molecular basis of this translocation, we calculated a three-dimensional map of the fully loaded (PA63)7-(LF)3 prepore complex by cryo-electron microscopy (cryo-EM). The map shows three LFs bound in a similar way to one another, via their N-terminal domains, to the surface of the PA heptamer. The model also reveals contacts between the N- and C-terminal domains of adjacent LF molecules. We propose that this molecular arrangement plays an important role in the maintenance of translocation efficiency through the narrow PA pore.
保护性抗原(PA)、致死因子(LF)和水肿因子(EF)。PA在寡聚化并形成前孔之前必须被宿主蛋白酶切割,LF和EF会结合到该前孔上。这种三方复合物被内吞后,前孔会转变为一个狭窄的跨膜孔,将未折叠的LF和EF递送到宿主细胞质中。在这里,我们发现多个90-kD的LF分子的转位迅速且高效。为了探究这种转位的分子基础,我们通过冷冻电子显微镜(cryo-EM)计算出了完全负载的(PA63)7-(LF)3前孔复合物的三维图谱。该图谱显示,三个LF通过它们的N端结构域以相似的方式结合到PA七聚体的表面。该模型还揭示了相邻LF分子的N端和C端结构域之间的接触。我们认为这种分子排列在通过狭窄的PA孔维持转位效率方面起着重要作用。