Rapid microsphere‐assisted peptide screening (MAPS) of promiscuous MHCII‐binding peptides in Zika virus envelope protein
Rapid microsphere‐assisted peptide screening (MAPS) of promiscuous MHCII‐binding peptides in Zika virus envelope protein
复制标题
寨卡病毒包膜蛋白中混杂的 MHCII 结合肽的快速微球辅助肽筛选 (MAPS)
DOI：
10.1002/aic.16697
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发表时间：
2019
期刊：
影响因子：
3.7
通讯作者：
Wen, Fei
中科院分区：
文献类型：
--
作者：
Smith, Mason R.;Bugada, Luke F.;Wen, Fei
尽管在计算工具方面取得了令人鼓舞的进展，但肽II类MHC（MHCII）结合预测因子仍然落后于肽I类MHC对应物。因此，肽-MHCII结合经常使用竞争性结合测定法进行实验评估，其倾向于牺牲通量以获得定量结合细节。在这里，我们开发了一种高通量半定量肽MHCII筛选策略，称为微球辅助肽筛选（MAPS），旨在平衡竞争性结合测定的准确性与计算工具的通量。使用MAPS，我们筛选了来自寨卡病毒包膜（E）蛋白的肽文库，用于结合四种常见的MHCII等位基因（DR 1、DR 4、DR 7、DR 15）。有趣的是，MAPS揭示了混杂结合多个MHCII等位基因和抗体中和位点的肽之间的显著重叠。在轮状病毒外壳糖蛋白VP 7中也观察到这种重叠，表明B细胞和CD 4 +T细胞特异性之间存在更深的关系，这有助于设计针对寨卡病毒和其他病毒的广泛保护性疫苗。
Despite promising developments in computational tools, peptide‐class II MHC (MHCII) binding predictors continue to lag behind their peptide‐class I MHC counterparts. Consequently, peptide–MHCII binding is often evaluated experimentally using competitive binding assays, which tend to sacrifice throughput for quantitative binding detail. Here, we developed a high‐throughput semiquantitative peptide–MHCII screening strategy termed microsphere‐assisted peptide screening (MAPS) that aims to balance the accuracy of competitive binding assays with the throughput of computational tools. Using MAPS, we screened a peptide library from Zika virus envelope (E) protein for binding to four common MHCII alleles (DR1, DR4, DR7, DR15). Interestingly, MAPS revealed a significant overlap between peptides that promiscuously bind multiple MHCII alleles and antibody neutralization sites. This overlap was also observed for rotavirus outer capsid glycoprotein VP7, suggesting a deeper relationship between B cell and CD4+T cell specificity which can facilitate the design of broadly protective vaccines to Zika and other viruses.