Eimeria tenella pyrroline -5-carboxylate reductase is a secreted protein and involved in host cell invasion

IF 1.4 4区医学 Q3 PARASITOLOGY

Experimental parasitology Pub Date : 2024-02-07 DOI:10.1016/j.exppara.2024.108712

Shanshan Liang , Shunhai Zhu , Qingjie Wang , Qiping Zhao , Hui Dong , Bing Huang , Yu Yu , Hongyu Han

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Abstract

Chicken coccidiosis, which caused by Eimeria spp, is a parasitic protozoal disease. At present, control measures of this disease depend mainly on anticoccidial drugs and live vaccines. But these control strategies have drawbacks such as drug resistance and limitations in live vaccines production. Therefore, novel control approaches are urgently need to study to control this disease effectively. In this study, the function and characteristics of the pyrroline-5-carboxylate reductase of Eimeria tenella (EtPYCR) protein were preliminary analyzed. The transcription and translation level were analyzed by using qPCR and Western blot. The results showed that the mRNA transcription and translation levels of EtPYCR were higher in unsporulated oocysts (UO) and second generation merozoites (Mrz) than that in sporulated oocysts (SO) and sporozoites. Enzyme activity showed that the enzyme activity of EtPYCR was also higher in the UO and Mrz than that in the SO and sporozoites. Immunofluorescence localization showed EtPYCR was mainly located on the top of sporozoites and the whole cytoplasm and surface of Mrz. The secretion assay indicated that EtPYCR was secretion protein, but not from micronemes. Invasion inhibition assay showed that rabbit anti-rEtPYCR polyclonal antibodies can effectively inhibit sporozoite invasion of DF-1 cells. These results showed that EtPYCR possess several important roles that separate and distinct from its conversion 1-pyrroline-5-carboxylate (P5C) into proline and maybe involved in the host cell invasion and development of parasites in host cells.

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天牛埃默氏菌吡咯啉-5-羧酸还原酶是一种分泌蛋白，参与宿主细胞的侵袭。

由艾美耳菌属引起的鸡球虫病是一种寄生性原虫病。目前，该疾病的控制措施主要依靠抗球虫药物和活疫苗。但这些控制策略存在抗药性和活疫苗生产局限性等缺点。因此，迫切需要研究新的控制方法来有效控制该疾病。本研究初步分析了天牛埃默氏菌吡咯啉-5-羧酸还原酶（EtPYCR）蛋白的功能和特性。采用 qPCR 和 Western 印迹技术分析了其转录和翻译水平。结果表明，未孢子化卵囊（UO）和第二代子囊虫（Mrz）中 EtPYCR 的 mRNA 转录和翻译水平均高于孢子化卵囊（SO）和子囊虫。酶活性显示，未孢子化卵囊（UO）和第二代裂殖体（Mrz）中的 EtPYCR 酶活性也高于 SO 和孢子虫中的 EtPYCR 酶活性。免疫荧光定位显示，EtPYCR主要位于孢子虫的顶部和Mrz的整个细胞质和表面。分泌试验表明，EtPYCR是一种分泌蛋白，但不是来自微粒体。侵袭抑制实验表明，兔抗EtPYCR多克隆抗体能有效抑制孢子虫对DF-1细胞的侵袭。这些结果表明，EtPYCR具有几个不同于其将1-吡咯啉-5-羧酸（P5C）转化为脯氨酸的重要作用，并可能参与宿主细胞的侵袭和寄生虫在宿主细胞中的发育。

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来源期刊

Experimental parasitology 医学-寄生虫学

CiteScore

3.10

自引率

4.80%

发文量

160

审稿时长

3 months

期刊介绍： Experimental Parasitology emphasizes modern approaches to parasitology, including molecular biology and immunology. The journal features original research papers on the physiological, metabolic, immunologic, biochemical, nutritional, and chemotherapeutic aspects of parasites and host-parasite relationships.