尖吻蝮蛇毒腺的缓激肽2型受体结合多肽的筛选与验证  

Screening and Validation of Binding Peptide of Type 2 Bradykinin Receptor from Venomous Gland of Deinagkistrodon acutus

作　　者：蒋安娜 刘杨 刘桂娜 唐业忠[2] 陆方[1] JIANG Anna;LIU Yang;LIU Guina;TANG Yezhong;LU Fang(West China Hospital,Sichuan University,Chengdu 610041,China;Chengdu Institute of Biology,Chinese Academy of Sciences,Chengdu 610041,China)

机构地区：[1]四川大学华西医院,成都610041 [2]中国科学院成都生物研究所,成都610041

出　　处：《四川动物》2024年第4期361-372,共12页Sichuan Journal of Zoology

摘　　要：蛇毒已被证明具有抗炎活性,其中包含许多免疫原性弱、活性强的短肽小分子。蛇毒腺是分泌毒液的器官,含有毒素成分及调控其分泌过程的大量生物活性成分。缓激肽2型受体(B2R)参与重要的细胞内信号通路,作为炎症的调节因子成为潜在的抗炎药物开发的靶点。为了获得特异性的B2R结合肽,利用尖吻蝮蛇Deinagkistro-don acutus毒腺T7噬菌体文库对靶点B2R进行3轮生物淘选,测序获得了多个具有潜在结合能力的序列。通过序列长度、溶解性预测、稳定性预测和分子对接等一系列生物信息学分析,最终确定1个含25个氨基酸的肽段,命名为DAvp-4。合成该肽段,通过表面等离子体共振技术验证了DAvp-4和B2R的结合能力,在脂多糖诱导的巨噬细胞模型及糖尿病视网膜病变小鼠模型中肯定了其抗炎作用。此研究不但获得了一个具有成药潜力的毒腺多肽,还显示了噬菌体展示技术在筛选天然产物成分研究中的有效性。Snake venomous glands contain many small peptides with weak immunogenicity and high biological activities including anti-inflammatory activity.The type 2 bradykinin receptor(B2R)is involved in several important intracellular signaling pathways and is considered as a potential target for anti-inflammatory drug development.In order to obtain specific B2R binding peptides for drug screening,the T7 phage library of Deinagkistrodon acutus venomous gland was used to perform three rounds of biological screening with B2R as the target.Multiple sequences with potential binding ability were obtained.Through a series of bioinformatic analyses such as sequence length,solubility predicting,stability predicting,and molecular docking,a 25 amino acids peptide segment,named DAvp-4,was ultimately determined.The peptide was synthesized,and then the binding ability of DAvp-4 to B2R was measured by surface plasma resonance.The antiinflammatory effect of DAvp-4 was confirmed in both macrophage cells injured with lipopolysaccharide and diabetic retinopathy mouse models.This study obtains a venomous gland peptide with patent medicine potential,but also demonstrates the effectiveness of phage display technology in screening natural product components.

关 键 词：噬菌体展示 毒腺 表面等离子体共振 缓激肽2型受体 

分 类 号：Q74[生物学—分子生物学] Q959.6