The Plasmodium falciparum exported protein PF3D7_0402000 binds to erythrocyte ankyrin and band 4.1.
The Plasmodium falciparum exported protein PF3D7_0402000 binds to erythrocyte ankyrin and band 4.1.
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DOI：
10.1016/j.molbiopara.2017.06.002
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发表时间：
2017-09
影响因子：
1.5
通讯作者：
LaCount DJ
中科院分区：
文献类型：
--
作者：
Shakya B;Penn WD;Nakayasu ES;LaCount DJ
恶性疟原虫广泛改变感染的红细胞（RBC），导致变形能力，形状和表面性质的变化。这些改变表明，红细胞细胞骨架是感染过程中修饰的主要目标。然而，导致这些变化的分子机制在很大程度上是未知的。为了开始解决这个问题，我们筛选了与红细胞细胞骨架蛋白锚蛋白1（ANK 1）和条带4.1（4.1R）结合的输出恶性疟原虫蛋白，所述锚蛋白1和条带4.1与有助于RBC的变形性和稳定性的其他细胞骨架蛋白形成关键的相互作用。用ANK 1和4.1R进行的酵母双杂交筛选鉴定了与恶性疟原虫输出蛋白的8种相互作用，包括4.1R和PF3D7_0402000（PFD 0090 c）之间的相互作用。这种相互作用首先在大规模筛选中鉴定（Vignali et al.，Malaria J，7：211，2008），其也报道了PF3D7_0402000和ANK 1之间的相互作用。我们在成对酵母双杂交和共沉淀测定中证实了PF3D7_0402000与4.1R和ANK 1的相互作用。在这两种情况下，需要PF3D7_0402000中的完整PHIST结构域进行结合。复合物纯化后的质谱分析为PF3D7_0402000与ANK 1和4.1R的相互作用提供了额外的支持。装载麦芽糖结合蛋白（MBP）-PF3D7_0402000的RBC血影细胞通过金属微球柱的效率低于装载模拟物或MBP的对照，这与PF3D7_0402000对RBC刚性或膜稳定性的影响一致。本研究在多项独立试验中证实了PF3D7_0402000与4.1R的相互作用，首次提供了PF3D7_0402000也与ANK 1结合的证据，并表明PF3D7_0402000影响未感染RBC血影的变形性或膜稳定性。
Plasmodium falciparum extensively modifies the infected red blood cell (RBC), resulting in changes in deformability, shape and surface properties. These alterations suggest that the RBC cytoskeleton is a major target for modification during infection. However, the molecular mechanisms leading to these changes are largely unknown. To begin to address this question, we screened for exported P. falciparum proteins that bound to the erythrocyte cytoskeleton proteins ankyrin 1 (ANK1) and band 4.1 (4.1R), which form critical interactions with other cytoskeletal proteins that contribute to the deformability and stability of RBCs. Yeast two-hybrid screens with ANK1 and 4.1R identified eight interactions with P. falciparum exported proteins, including an interaction between 4.1R and PF3D7_0402000 (PFD0090c). This interaction was first identified in a large-scale screen (Vignali et al., Malaria J, 7:211, 2008), which also reported an interaction between PF3D7_0402000 and ANK1. We confirmed the interactions of PF3D7_0402000 with 4.1R and ANK1 in pair-wise yeast two-hybrid and co-precipitation assays. In both cases, an intact PHIST domain in PF3D7_0402000 was required for binding. Complex purification followed by mass spectrometry analysis provided additional support for the interaction of PF3D7_0402000 with ANK1 and 4.1R. RBC ghost cells loaded with maltose-binding protein (MBP)-PF3D7_0402000 passed through a metal microsphere column less efficiently than mock- or MBP-loaded controls, consistent with an effect of PF3D7_0402000 on RBC rigidity or membrane stability. This study confirmed the interaction of PF3D7_0402000 with 4.1R in multiple independent assays, provided the first evidence that PF3D7_0402000 also binds to ANK1, and suggested that PF3D7_0402000 affects deformability or membrane stability of uninfected RBC ghosts.
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影响因子：
64.8
作者：
LaCount, DJ;Vignali, M;Hughes, RE
通讯作者：
Hughes, RE
影响因子：
20.3
作者：
Anong, William A.;Franco, Taina;Low, Philip S.
通讯作者：
Low, Philip S.
影响因子：
4.4
作者：
Frech C;Chen N
通讯作者：
Chen N
影响因子：
1.5
作者：
Brown, Hakeenah F.;Wang, Ling;LaCount, Douglas J.
通讯作者：
LaCount, Douglas J.
影响因子：
2.9
作者：
Ipsaro, Jonathan J.;Huang, Lei;MacDonald, Ruby I.
通讯作者：
MacDonald, Ruby I.