Specific EtMIC3-binding peptides inhibit Eimeria tenella sporozoites entry into host cells.
Specific EtMIC3-binding peptides inhibit Eimeria tenella sporozoites entry into host cells.
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DOI：
10.1186/s13567-020-00873-y
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发表时间：
2021-02-17
影响因子：
4.4
通讯作者：
Ma D
中科院分区：
文献类型：
--
作者：
Chen W;Ma C;Li G;Jia Z;Yang X;Pan X;Ma D
艾美耳球虫引起的禽球虫病给全球养禽业造成了巨大的经济损失。本研究以柔嫩艾美耳球虫微丝蛋白3(EtMIC3-Bc1)的微丝黏附重复区(MARR)Bc1为配体，从噬菌体展示肽库中筛选与EtMIC3结合的多肽。用酶联免疫吸附试验(EL ISA)检测阳性噬菌体克隆。用竞争酶联免疫吸附试验进一步验证阳性噬菌体与重组EtMIC3-bc1蛋白或子孢子蛋白的结合能力。体外检测目的多肽对子孢子侵袭MDBK细胞的抑制作用。给鸡口服靶向阳性噬菌体，评价其对同源攻击的保护作用。建立了EtMIC3-Bc1的三维结构模型，分析了目标多肽与EtMIC3-Bc1模型之间的分子对接。结果表明，筛选出的3个阳性噬菌体均能与EtMIC3-Bc1蛋白特异性结合。3种多肽A、D和W均能有效抑制MDBK细胞由子孢子侵袭，抑制率分别为71.8%、54.6%和20.8%。口服噬菌体A组的鸡对同源攻击的保护力明显高于其他组。分子对接表明，三种多肽中的氨基酸，尤其是A肽中的氨基酸插入到EtMIC3-Bc1蛋白的疏水槽中，并通过分子间氢键与EtMIC3-Bc1结合。综上所述，结果表明EtMIC3结合肽抑制子孢子进入宿主细胞。本研究为探索防治球虫病的新策略提供了新的思路。
Avian coccidiosis caused by Eimeria leads to huge economic losses on the global poultry industry. In this study, microneme adhesive repeat regions (MARR) bc1 of E. tenella microneme protein 3 (EtMIC3-bc1) was used as ligand, and peptides binding to EtMIC3 were screened from a phage display peptide library. The positive phage clones were checked by enzyme-linked immunosorbent assay (ELISA). Competitive ELISA was applied to further verify the binding capability between the positive phages and recombinant EtMIC3-bc1 protein or sporozoites protein. The inhibitory effects of target peptides on sporozoites invasion of MDBK cells were measured in vitro. Chickens were orally administrated with target positive phages and the protective effects against homologous challenge were evaluated. The model of three-dimensional (3D) structure for EtMIC3-bc1 was conducted, and molecular docking between target peptides and EtMIC3-bc1 model was analyzed. The results demonstrated that three selected positive phages specifically bind to EtMIC3-bc1 protein. The three peptides A, D and W effectively inhibited invasion of MDBK cells by sporozoites, showing inhibited ratio of 71.8%, 54.6% and 20.8%, respectively. Chickens in the group orally inoculated with phages A displayed more protective efficacies against homologous challenge than other groups. Molecular docking showed that amino acids in three peptides, especially in peptide A, insert into the hydrophobic groove of EtMIC3-bc1 protein, and bind to EtMIC3-bc1 through intermolecular hydrogen bonds. Taken together, the results suggest EtMIC3-binding peptides inhibit sporozoites entry into host cells. This study provides new idea for exploring novel strategies against coccidiosis.
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影响因子：
1.5
作者：
Labbé, M;de Venevelles, P;Péry, P
通讯作者：
Péry, P
影响因子：
4.8
作者：
Periz, Javier;Gill, Andrew C.;Tomley, Fiona M.
通讯作者：
Tomley, Fiona M.
影响因子：
2
作者：
Periz, Javier;Ryan, Rachael;Tomley, Fiona M.
通讯作者：
Tomley, Fiona M.
影响因子：
6.7
作者：
Lai L;Bumstead J;Liu Y;Garnett J;Campanero-Rhodes MA;Blake DP;Palma AS;Chai W;Ferguson DJ;Simpson P;Feizi T;Tomley FM;Matthews S
通讯作者：
Matthews S
影响因子：
6.7
作者：
Collins, Christine R.;Withers-Martinez, Chrislaine;Hackett, Fiona;Blackman, Michael J.
通讯作者：
Blackman, Michael J.