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本研究首次报道具有牙釉质结合活性的环肽序列(-CMPQVMPMC-)。在反筛试验中,投入等量牙釉质粉末和相同滴度噬菌体,外壳蛋白pⅢ上表达该展示肽的噬菌体回收量明显高于不含展示肽的野生型噬菌体M13。固相法体外合成该环状七肽以模拟展示肽在噬菌体表面的构象,并利用96孔板体外快速凝胶单扩散系统验证其对钙/磷沉积的影响。矿化开始后的24h内,每隔6h记录各样本在405nm的OD值。各样本的相对浊度表示为所占24h末空白组OD值增量(100%)的百分比。结果显示,牙釉质结合肽(EBP)能明显延迟凝胶中的钙磷成核,且延迟效果与添加的浓度相关。在矿化24h末,含结合肽的各组凝胶内浊度明显低于空白组,但高于BSA组。
This study for the first time reported the enamel binding activity of cyclic peptide sequence (-CMPQVMPMC-). In the reverse sieve test, the same amount of enamel powder and the same titer of phage were injected, and the amount of phage displayed on the coat protein pIII was significantly higher than that of the wild-type phage M13 without peptide. The cyclic heptapeptide was synthesized in vitro by a solid-phase method to simulate the conformation of the peptide on the phage surface and the effect of calcium / phosphorus deposition on the phage surface was verified by a 96-well plate in vitro rapid gel diffusion system. Within 24 h after the start of mineralization, OD values of each sample at 405 nm were recorded every 6 h. The relative turbidity of each sample is expressed as a percentage of the increase (100%) in the OD value of the blank group at the end of 24 h. The results showed that enamel binding peptide (EBP) significantly delayed calcium and phosphorus nucleation in the gel, and the delay effect was correlated with the added concentration. At the end of 24 h of mineralization, the gels in the gel-containing groups were significantly lower than the blank group, but higher than the BSA group.