噬菌体展示技术筛选抗轮状病毒多肽的试验研究

Screening for Peptides of Anti-rotavirus by Phage-displayed Technique

摘要:

从噬菌体随机展示十五肽文库筛选出4个与轮状病毒粒子特异性结合多肽。经空斑减少抑制实验和MTT法分析表明其中3个多肽对病毒感染培养细胞具有抑制作用，其中序列为QSNPIHIITNTRNHP的C肽具有显著抑制作用，抑制效果达93%，另外2个多肽A和B抑制效果分别为40%与50%。经过多肽序列分析发现这3个十五肽具有2个保守序列，分别是第2至8个氨基酸残基SNPIHII和第12～15个氨基酸残基NIP。胰蛋白酶水解位点分析表明C肽无裂解位点，而A肽和B肽则分别具有3个和4个潜在水解位点。抑制病毒感染液中胰蛋白酶活性，发现A，B两肽也能显著地抑制病毒离体感染。说明所筛选的多肽2个保守序列的完整对抗病毒感染起着重要作用。C肽有望成为一种治疗轮状病毒感染的口服药物。

Abstract:

In this study，a 15-mer phage display peptide library was employed to pan against human rotavirus immobilized on solid phase. 4 different peptides were selected and could bind with rotavirus particles specifically. Plaque reduction neutralization test and MTT analysis results indicated that 3 of the peptides can inhibit rotavirus infecting in vitro. A peptide which sequence is QSNPIHIITNTRNHP showed the best efficiency—93% neutralization infectivity. Two other peptides，A and B，showed 40% and 50% neutralization infectivity respectively. Amino sequence analysis results indicate the 3 peptides containing 2 conserved motifs: SNPIHII and NIP. No putative trypsin hydrolysis site was found in C peptide，however，4 and 3 potential sites were found in A and B peptides respectively. Using trypsin inhibitor，both A and B peptides showed the similar antiviral effect as that of C peptide. It suggests that the intactness of the 2 conserved motifs play an important role in counteracting virus infection. According to the results of this study，peptide C is hopeful to be exploited as an antiviral peptide drug.