A member of the Plasmodium falciparum PHIST family binds to the erythrocyte cytoskeleton component band 4.1.
A member of the Plasmodium falciparum PHIST family binds to the erythrocyte cytoskeleton component band 4.1.
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DOI：
10.1186/1475-2875-12-160
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发表时间：
2013-05-11
期刊：
影响因子：
3
通讯作者：
Rayner JC
中科院分区：
文献类型：
--
作者：
Parish LA;Mai DW;Jones ML;Kitson EL;Rayner JC
恶性疟原虫寄生虫将400多种蛋白质输出到其宿主红细胞的胞质溶胶中。这些输出的蛋白质催化红细胞质膜上的结节的形成和红细胞刚性的总体增加，推测是通过调节内源性红细胞细胞骨架。在未感染的红细胞中，带4.1（4.1R）通过其三叶草形N-末端结构域的三个叶与多种蛋白质相互作用，在调节红细胞形状中起关键作用。在恶性疟原虫感染的红细胞中，4.1R的C-叶与恶性疟原虫蛋白成熟寄生虫感染的红细胞表面抗原（梅萨）相互作用，但目前尚不清楚其他恶性疟原虫蛋白是否与4.1R N-末端结构域的其他叶结合。为了鉴定新的4.1R相互作用蛋白，用含有N-和α-叶的4.1R片段进行酵母双杂交筛选。使用定点诱变证实并研究了阳性相互作用，并针对相互作用伴侣产生抗体以抑制其在恶性疟原虫感染的红细胞中的表达和分布。酵母双杂交筛选鉴定了4.1R N-和α-叶与PF3D7_0402000之间的正相互作用。PF3D7_0402000是一个大家族的输出蛋白，共享一个未知功能的结构域，PHIST结构域的成员。结构域定位和定点突变确定了与4.1R相互作用的是PF3D7_0402000的PHIST结构域。天然PF3D7_0402000定位于寄生虫液泡膜（PVM），并与4.1R亚群共定位。大多数恶性疟原虫输出蛋白（包括PHIST家族的大多数成员）的功能尚不清楚，仅在少数情况下，恶性疟原虫输出蛋白与红细胞细胞骨架组分之间存在直接相互作用。4.1R和PF3D7_0402000之间的相互作用，以及PF3D7_0402000在PVM上的4.1R亚群的定位可能表明在调节PVM结构中的作用。需要进一步研究4.1R招募的机制。PF3D7_0402000被鉴定为主要红细胞骨架蛋白4.1R的新结合伴侣。这种相互作用与越来越多的文献一致，这些文献表明PHIST家族成员通过直接与红细胞蛋白相互作用发挥作用。
Plasmodium falciparum parasites export more than 400 proteins into the cytosol of their host erythrocytes. These exported proteins catalyse the formation of knobs on the erythrocyte plasma membrane and an overall increase in erythrocyte rigidity, presumably by modulating the endogenous erythrocyte cytoskeleton. In uninfected erythrocytes, Band 4.1 (4.1R) plays a key role in regulating erythrocyte shape by interacting with multiple proteins through the three lobes of its cloverleaf-shaped N-terminal domain. In P. falciparum-infected erythrocytes, the C-lobe of 4.1R interacts with the P. falciparum protein mature parasite-infected erythrocyte surface antigen (MESA), but it is not currently known whether other P. falciparum proteins bind to other lobes of the 4.1R N-terminal domain. In order to identify novel 4.1R interacting proteins, a yeast two-hybrid screen was performed with a fragment of 4.1R containing both the N- and α-lobes. Positive interactions were confirmed and investigated using site-directed mutagenesis, and antibodies were raised against the interacting partner to characterise it’s expression and distribution in P. falciparum infected erythrocytes. Yeast two-hybrid screening identified a positive interaction between the 4.1R N- and α-lobes and PF3D7_0402000. PF3D7_0402000 is a member of a large family of exported proteins that share a domain of unknown function, the PHIST domain. Domain mapping and site-directed mutagenesis established that it is the PHIST domain of PF3D7_0402000 that interacts with 4.1R. Native PF3D7_0402000 is localized at the parasitophorous vacuole membrane (PVM), and colocalizes with a subpopulation of 4.1R. The function of the majority of P. falciparum exported proteins, including most members of the PHIST family, is unknown, and in only a handful of cases has a direct interaction between P. falciparum-exported proteins and components of the erythrocyte cytoskeleton been established. The interaction between 4.1R and PF3D7_0402000, and localization of PF3D7_0402000 with a sub-population of 4.1R at the PVM could indicate a role in modulating PVM structure. Further investigation into the mechanisms for 4.1R recruitment is needed. PF3D7_0402000 was identified as a new binding partner for the major erythrocyte cytoskeletal protein, 4.1R. This interaction is consistent with a growing body of literature that suggests the PHIST family members function by interacting directly with erythrocyte proteins.
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影响因子：
9.8
作者：
Bozdech, Zbynek;Llinas, Manuel;Pulliam, Brian Lee;Wong, Edith D;Zhu, Jingchun;DeRisi, Joseph L
通讯作者：
DeRisi, Joseph L
影响因子：
4.8
作者：
Manno, S;Takakuwa, Y;Mohandas, N
通讯作者：
Mohandas, N
DOI：
10.1074/jbc.m111.330779
发表时间：
2012-03-02
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Mayer C;Slater L;Erat MC;Konrat R;Vakonakis I
通讯作者：
Vakonakis I
影响因子：
4.8
作者：
Miller, SK;Good, RT;Crabb, BS
通讯作者：
Crabb, BS
影响因子：
5.3
作者：
Spycher, Cornelia;Rug, Melanie;Tilley, Leann
通讯作者：
Tilley, Leann