Characterization and utility of phages bearing peptides with affinity to porcine reproductive and respiratory syndrome virus nsp7 protein
Characterization and utility of phages bearing peptides with affinity to porcine reproductive and respiratory syndrome virus nsp7 protein
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携带对猪繁殖与呼吸综合征病毒 nsp7 蛋白有亲和力的肽的噬菌体的表征和用途
DOI：
10.1016/j.jviromet.2015.04.026
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发表时间：
2015-09-15
影响因子：
3.1
通讯作者：
Zhang, Guihong
中科院分区：
文献类型：
--
作者：
Wang, Heng;Liu, Rongchang;Zhang, Guihong
利用噬菌体展示技术筛选出与猪繁殖与呼吸综合征病毒（PRRSV）非结构蛋白（nsp）7具有高亲和力的多肽。经过6轮生物淘选，鉴定出5个12个氨基酸的肽序列。一个推定的CD##WC基序被发现在两个不同的共识肽所承担的E14和5。根据结合测定的结果，合成由α 4、5和6携带的肽用于随后的实验。免疫荧光实验表明，这些肽均能识别PRRSV感染细胞中的nsp7。此外，这些肽显示出抗病毒活性，其中肽5和6显示出有效的抑制作用。早期肽刺激与强的抗病毒活性相关，并且肽的抑制作用在感染后36和48 h具有剂量依赖性。选择肽5通过共聚焦显微镜检测nsp7的细胞内定位。该肽对nsp7具有与抗nsp7单克隆抗体相似的作用。这些结果表明，对PRRSV nsp7的高亲和力肽可以模拟nsp7抗体作为用于病毒检测的诊断试剂的潜力。此外，本研究中选择的肽代表了抑制PRRSV的潜在有效的抗病毒候选物。(C)2015年由Elsevier B.V.出版
High-affinity peptides to porcine reproductive and respiratory syndrome virus (PRRSV) nonstructural protein (nsp) 7 were identified using phage-display technology. Five 12-amino-acid peptide sequences were identified after six rounds of biopanning. A putative CD##WC motif was found in two different consensus peptides borne by phages 4 and 5. The peptides borne by phages 4, 5, and 6 were synthesized for subsequent experiments, according to the results of the binding assays. Immunofluorescence assay revealed that all these peptides recognized nsp7 in PRRSV-infected cells. Furthermore, the peptides demonstrated antiviral activities, with peptides 5 and 6 showing effective inhibition. Early peptide stimulation was associated with strong antiviral activity, and the inhibitory effects of the peptides were dose-dependent at 36 and 48 h post-infection. Peptide 5 was selected to detect the intracellular localization of nsp7 by confocal microscopy. This peptide had a similar effect to anti-nsp7 monoclonal antibody on nsp7. These results suggest that high-affinity peptides to PRRSV nsp7 could mimic the potential of nsp7 antibody as a diagnostic reagent for virus detection. Moreover, the peptides selected in this study represented a potentially effective antiviral candidate to inhibit PRRSV. (C) 2015 Published by Elsevier B.V.