应用噬菌体展示肽库技术淘选大鼠CCR5膜外第一、二胞外环特异性结合的活性拮抗肽与初步鉴定

发布时间：2018-04-02 23:35

本文选题：CC趋化因子受体　切入点：拮抗肽　出处：《中国病理生理杂志》2015年07期

【摘要】：目的:利用噬菌体展示肽库技术淘选与大鼠CC趋化因子受体5(CCR5)膜外第一、二胞外环特异性结合的短肽,并鉴定其与CCR5的结合能力。方法:在蛋白质数据库中查得大鼠CCR5第一、二胞外环的氨基酸序列,合成相应的线性短肽作为淘选的靶分子,利用噬菌体展示7肽文库进行3~4轮淘选,用ELISA法鉴定所选肽与靶分子的结合,并测定其与浓度的关系。结果:与CCR5第一、二胞外环特异性结合的噬菌体展示的短肽序列分别为GHWKVWL和HYIDFRW,ELISA鉴定呈阳性反应,且短肽与靶分子的结合具有浓度依赖性和可饱和性。结论:利用噬菌体展示技术成功获得了2条CCR5特异性结合的短肽,并在体外证明其可与CCR5第一、二胞外环具有结合能力。
[Abstract]:Aim: to elucidate the specific binding of short peptides to rat CC chemokine receptor 5 (CCR5) by phage display peptide library, and to identify the binding ability of the peptide to CCR5.Methods: the amino acid sequences of the first and second extracellular rings of rat CCR5 were identified in the protein database, and the corresponding linear short peptides were synthesized as target molecules for panning. The phage display 7 peptide library was used for 34 rounds of panning.The binding of the selected peptide to the target molecule was identified by ELISA, and the relationship between the peptide and the concentration was determined.Results: the short peptide sequences displayed by phage binding to the first and second extracellular rings of CCR5 were GHWKVWL and HYIDFRWN Elisa, respectively, and the binding of short peptides to target molecules was concentration-dependent and saturable.Conclusion: two short peptides of CCR5 binding were successfully obtained by phage display technique, and it was proved in vitro that they could bind to the first and second extracellular rings of CCR5.
【作者单位】：中山大学孙逸仙纪念医院消化内科;广东药学院附属第一医院消化内科;中山大学孙逸仙纪念医院儿科;
【基金】：国家自然科学资金基助项目(No.81370499) 广东省自然科学基金资助项目(No.2014-A030313020)
【分类号】：R574.62

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