Analysis of the interactome of the Ser/Thr Protein Phosphatase type 1 in Plasmodium falciparum.
Analysis of the interactome of the Ser/Thr Protein Phosphatase type 1 in Plasmodium falciparum.
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DOI：
10.1186/s12864-016-2571-z
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发表时间：
2016-03-17
期刊：
影响因子：
4.4
通讯作者：
Khalife J
中科院分区：
文献类型：
--
作者：
Hollin T;De Witte C;Lenne A;Pierrot C;Khalife J
蛋白磷酸酶1 （PP1）是疟原虫恶性疟原虫（Pf）细胞生存所必需的酶。PP1的活性受调节亚基结合的调节，人类中有多达200个调节亚基，但迄今为止仅报道了3个用于寄生虫。为了更好地了解恶性疟原虫PP1 （PfPP1）调控网络，我们在这里报告了使用三种策略来表征PfPP1相互作用组：通过质谱鉴定的共亲和纯化蛋白，酵母双杂交（Y2H）筛选和恶性疟原虫预测蛋白质组的计算机分析。共亲和纯化和质谱分析鉴定了6个PfPP1相互作用蛋白（Pips），其中3个包含RVxF一致结合，2个具有Fxx[RK]x[RK]基序，也被证明是PP1结合基序，一个具有两个结合基序。Y2H筛选鉴定出134个蛋白，其中30个具有RVxF结合基序，20个具有Fxx[RK]x[RK]结合基序。使用共识RVxF基序作为模板的Pf预测蛋白质组的计算机屏幕显示存在55个潜在的pip。作为进一步的证明，35个候选蛋白在基于elisa的分析中被验证为PfPP1相互作用蛋白。据我们所知，这是对PfPP1相互作用组的首次研究。这些数据报告了几个保守的PP1相互作用蛋白以及大量的PfPP1特异性相互作用蛋白。他们的分析表明，PP1在疟原虫中具有高度多样性的生物学功能。基于目前的数据和对Pf相互作用组的早期研究，提出了Pips在蛋白质折叠/蛋白质水解、转录和致病性网络中的潜在含义。目前的工作为进一步研究这些相互作用的结构基础及其在恶性疟原虫中的功能提供了一个起点。本文的在线版本（doi:10.1186/s12864-016-2571-z）包含补充材料，可供授权用户使用。
Protein Phosphatase 1 (PP1) is an enzyme essential to cell viability in the malaria parasite Plasmodium falciparum (Pf). The activity of PP1 is regulated by the binding of regulatory subunits, of which there are up to 200 in humans, but only 3 have been so far reported for the parasite. To better understand the P. falciparum PP1 (PfPP1) regulatory network, we here report the use of three strategies to characterize the PfPP1 interactome: co-affinity purified proteins identified by mass spectrometry, yeast two-hybrid (Y2H) screening and in silico analysis of the P. falciparum predicted proteome. Co-affinity purification followed by MS analysis identified 6 PfPP1 interacting proteins (Pips) of which 3 contained the RVxF consensus binding, 2 with a Fxx[RK]x[RK] motif, also shown to be a PP1 binding motif and one with both binding motifs. The Y2H screens identified 134 proteins of which 30 present the RVxF binding motif and 20 have the Fxx[RK]x[RK] binding motif. The in silico screen of the Pf predicted proteome using a consensus RVxF motif as template revealed the presence of 55 potential Pips. As further demonstration, 35 candidate proteins were validated as PfPP1 interacting proteins in an ELISA-based assay. To the best of our knowledge, this is the first study on PfPP1 interactome. The data reports several conserved PP1 interacting proteins as well as a high number of specific interactors to PfPP1. Their analysis indicates a high diversity of biological functions for PP1 in Plasmodium. Based on the present data and on an earlier study of the Pf interactome, a potential implication of Pips in protein folding/proteolysis, transcription and pathogenicity networks is proposed. The present work provides a starting point for further studies on the structural basis of these interactions and their functions in P. falciparum. The online version of this article (doi:10.1186/s12864-016-2571-z) contains supplementary material, which is available to authorized users.