Characterization of Two EF-hand Domain-containing Proteins from Toxoplasma gondii.
Characterization of Two EF-hand Domain-containing Proteins from Toxoplasma gondii.
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DOI：
10.1111/jeu.12675
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发表时间：
2019-03
期刊：
影响因子：
--
通讯作者：
Hortua Triana MA
中科院分区：
文献类型：
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作者：
Chang L;Dykes EJ;Li J;Moreno SNJ;Hortua Triana MA
钙离子（Ca 2+）作为细胞内第二信使的普遍作用依赖于大量的Ca 2+结合蛋白（CBP），其能够通过特异性结构域结合Ca 2+。许多CBP共享一种称为EF-手的Ca 2+结合结构域。EF-手基序已被充分研究，并由一个螺旋-环-螺旋结构域与特定的氨基酸在环区域，与钙离子相互作用。在弓形虫的大量基因（约68）预测至少有一个EF-手形基序。这些基因中的大多数尚未被表征。我们报告了两种含有EF-手基序的蛋白质TgGT1_216620和TgGT1_280480的特征，它们分别定位于质膜和棒状球。通过CRISPR（成簇的规则间隔短回文重复序列）/Cas9（CRISPR相关蛋白9）对这些基因的遗传破坏导致突变寄生虫克隆（Δ tg 216620和Δ tg 280480）的生长速度低于对照细胞。Ca ~（2+）测定结果表明，Δ tg 216620细胞对细胞外Ca ~（2+）无反应，而Δ tg 280480细胞对细胞外Ca ~（2+）有反应。我们的假设是，TgGT1_216620是重要的Ca 2+内流，而TgGT1_280480可能发挥不同的作用，在棒状体。
The universal role of calcium (Ca2+) as a second messenger in cells depends on a large number of Ca2+-binding proteins (CBP), which are able to bind Ca2+ through specific domains. Many CBPs share a type of Ca2+-binding domain known as the EF-hand. The EF-hand motif has been well studied and consists of a helix-loop-helix structural domain with specific amino acids in the loop region that interact with Ca2+. In Toxoplasma gondii a large number of genes (approximately 68) are predicted to have at least one EF-hand motif. The majority of these genes have not been characterized. We report the characterization of two EF-hand motif-containing proteins, TgGT1_216620 and TgGT1_280480, which localize to the plasma membrane and to the rhoptiy bulb, respectively. Genetic disruption of these genes by CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9) resulted in mutant parasite clones (Δtg216620 and Δtg280480) that grew at a slower rate than control cells. Ca2+ measurements showed that Δtg216620 cells did not respond to extracellular Ca2+ as the parental controls while Δtg280480 cells appeared to respond as the parental cells. Our hypothesis is that TgGT1_216620 is important for Ca2+ influx while TgGT1_280480 may be playing a different role in the rhoptries.
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影响因子：
6.7
作者：
Sheiner L;Demerly JL;Poulsen N;Beatty WL;Lucas O;Behnke MS;White MW;Striepen B
通讯作者：
Striepen B
弓形虫的裂解周期：15年后。
影响因子：
10.5
作者：
Blader IJ;Coleman BI;Chen CT;Gubbels MJ
通讯作者：
Gubbels MJ
影响因子：
4.8
作者：
Pace, Douglas A.;McKnight, Ciara A.;Moreno, Silvia N. J.
通讯作者：
Moreno, Silvia N. J.
影响因子：
4
作者：
Arrizabalaga, G;Boothroyd, JC
通讯作者：
Boothroyd, JC
影响因子：
64.8
作者：
通讯作者：
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