Molecular characterization and analysis of a novel protein disulfide isomerase-like protein of Eimeria tenella.
Molecular characterization and analysis of a novel protein disulfide isomerase-like protein of Eimeria tenella.
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DOI：
10.1371/journal.pone.0099914
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Huang B
中科院分区：
文献类型：
--
作者：
Han H;Dong H;Zhu S;Zhao Q;Jiang L;Wang Y;Li L;Wu Y;Huang B
蛋白质二硫键异构酶（PDI）和PDI样蛋白是硫氧还蛋白超家族的成员。它们含有硫氧还蛋白样结构域，催化蛋白质二硫键的生理氧化、还原和异构化，参与原核生物和真核生物的细胞功能和发育。本研究根据表达序列标签（EST），利用cDNA末端快速扩增（RACE）技术克隆了柔嫩艾美耳球虫（Eimeria tenella）的一个新的类PDI基因EtPDIL。EtPDIL cDNA全长1129个核苷酸，编码216个氨基酸。EtPDIL蛋白属于硫氧还蛋白样超家族，具有一个硫氧还蛋白结构域，并具有一个非经典的硫氧还蛋白样基序（SXXC）。BLAST分析表明，EtPDIL蛋白与其他顶复门寄生虫的PDI样蛋白具有55-59%的相同性。采用实时荧光定量PCR和western blot检测EtPDIL在不同发育阶段的转录和蛋白水平。EtPDIL在孢子化卵囊中的mRNA和蛋白水平高于未孢子化卵囊、子孢子或裂殖子。在未形成孢子的卵囊中几乎检测不到蛋白质表达。Western印迹表明，兔抗血清对重组EtPDIL承认只有一个天然的24 kDa的蛋白质从寄生虫。EtPDIL抗体的免疫定位显示EtPDIL在完整的子孢子和裂殖子的胞质中呈分散分布。子孢子在完全培养基中孵育后，EtPDIL蛋白集中在子孢子的前部，并出现在寄生虫的表面。成熟疟原虫释放的裂殖子侵入DF-1细胞后，特异性染色更强，主要位于疟原虫表面。在DF-1细胞中寄生虫发育后，滋养体、未成熟滋养体和成熟滋养体中的染色增强。抗体抑制EtPDIL功能降低了E. tenella侵袭DF-1细胞。这些结果表明，EtPDIL可能参与了外环境中的孢子形成以及宿主细胞的粘附、侵袭和发育。tenella。
Protein disulfide isomerase (PDI) and PDI-like proteins are members of the thioredoxin superfamily. They contain thioredoxin-like domains and catalyze the physiological oxidation, reduction and isomerization of protein disulfide bonds, which are involved in cell function and development in prokaryotes and eukaryotes. In this study, EtPDIL, a novel PDI-like gene of Eimeria tenella, was cloned using rapid amplification of cDNA ends (RACE) according to the expressed sequence tag (EST). The EtPDIL cDNA contained 1129 nucleotides encoding 216 amino acids. The deduced EtPDIL protein belonged to thioredoxin-like superfamily and had a single predicted thioredoxin domain with a non-classical thioredoxin-like motif (SXXC). BLAST analysis showed that the EtPDIL protein was 55–59% identical to PDI-like proteins of other apicomplexan parasites. The transcript and protein levels of EtPDIL at different development stages were investigated by real-time quantitative PCR and western blot. The messenger RNA and protein levels of EtPDIL were higher in sporulated oocysts than in unsporulated oocysts, sporozoites or merozoites. Protein expression was barely detectable in unsporulated oocysts. Western blots showed that rabbit antiserum against recombinant EtPDIL recognized only a native 24 kDa protein from parasites. Immunolocalization with EtPDIL antibody showed that EtPDIL had a disperse distribution in the cytoplasm of whole sporozoites and merozoites. After sporozoites were incubated in complete medium, EtPDIL protein concentrated at the anterior of the sporozoites and appeared on the surface of parasites. Specific staining was more intense and mainly located on the parasite surface after merozoites released from mature schizonts invaded DF-1 cells. After development of parasites in DF-1 cells, staining intensified in trophozoites, immature schizonts and mature schizonts. Antibody inhibition of EtPDIL function reduced the ability of E. tenella to invade DF-1 cells. These results suggested that EtPDIL might be involved in sporulation in external environments and in host cell adhesion, invasion and development of E. tenella.
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影响因子：
2.9
作者：
Darby, NJ;Kemmink, J;Creighton, TE
通讯作者：
Creighton, TE
影响因子：
3.1
作者：
Ben Achour, Y;Chenik, M;Dellagi, K
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柔嫩艾美耳球虫新型钙依赖性蛋白激酶的分子表征和分析
影响因子：
2.4
作者：
Han, H. Y.;Zhu, S. H.;Huang, B.
通讯作者：
Huang, B.
影响因子：
1.5
作者：
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通讯作者：
Lipp, J
影响因子：
2
作者：
Jiang, Lianlian;Lin, Jiaojiao;Huang, Bing
通讯作者：
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