Jamwal Abhishek, Round Adam R, Bannwarth Ludovic, Venien-Bryan Catherine, Belrhali Hassan, Yogavel Manickam, Sharma Amit
From the Structural and Computational Biology Group, International Centre for Genetic Engineering and Biotechnology, New Delhi 110067, India.
the European Molecular Biology Laboratory, Grenoble Outstation, 38042 Grenoble, France, the Unit for Virus Host-Cell Interactions, University Grenoble Alpes-EMBL-CNRS, 38042 Grenoble, France, and.
J Biol Chem. 2015 Dec 18;290(51):30498-513. doi: 10.1074/jbc.M115.674176. Epub 2015 Oct 22.
Trypanosoma brucei (T. brucei) is responsible for the fatal human disease called African trypanosomiasis, or sleeping sickness. The causative parasite, Trypanosoma, encodes soluble versions of inorganic pyrophosphatases (PPase), also called vacuolar soluble proteins (VSPs), which are localized to its acidocalcisomes. The latter are acidic membrane-enclosed organelles rich in polyphosphate chains and divalent cations whose significance in these parasites remains unclear. We here report the crystal structure of T. brucei brucei acidocalcisomal PPases in a ternary complex with Mg(2+) and imidodiphosphate. The crystal structure reveals a novel structural architecture distinct from known class I PPases in its tetrameric oligomeric state in which a fused EF hand domain arranges around the catalytic PPase domain. This unprecedented assembly evident from TbbVSP1 crystal structure is further confirmed by SAXS and TEM data. SAXS data suggest structural flexibility in EF hand domains indicative of conformational plasticity within TbbVSP1.
布氏锥虫(T. brucei)是导致致命人类疾病——非洲锥虫病(又称昏睡病)的病原体。致病寄生虫锥虫编码无机焦磷酸酶(PPase)的可溶性形式,也称为液泡可溶性蛋白(VSP),这些蛋白定位于其酸性钙小体。酸性钙小体是富含多磷酸盐链和二价阳离子的酸性膜包被细胞器,其在这些寄生虫中的意义尚不清楚。我们在此报告布氏锥虫酸性钙小体PPase与Mg(2+)和亚氨基二磷酸形成的三元复合物的晶体结构。晶体结构揭示了一种新颖的结构架构,在其四聚体寡聚状态下与已知的I类PPase不同,其中一个融合的EF手结构域围绕催化PPase结构域排列。小角X射线散射(SAXS)和透射电子显微镜(TEM)数据进一步证实了从TbbVSP1晶体结构中明显看出的这种前所未有的组装。SAXS数据表明EF手结构域具有结构灵活性,这表明TbbVSP1内存在构象可塑性。