双靶向MMP-14和金属离子的结合肽筛选与分子模拟  

Selection and Molecular Simulation of Binding Peptides Dual-targeting MMP-14 and Metal Ions

作　　者：梁志娟[1] 黄敬双[1] 崔健[1] 熊莉丽[1] 茆灿泉[1] 

机构地区：[1]西南交通大学生命科学与工程学院分子进化与应用生物学实验室,成都610031

出　　处：《中国生物化学与分子生物学报》2011年第4期341-349,共9页Chinese Journal of Biochemistry and Molecular Biology

基　　金：中央高校基本科研业务费专项资金(No.SWJTU09ZT28);国家高科技研究发展计划(863计划;No.2006AAZ353)资助项目~~

摘　　要：以基质金属蛋白酶-14(MMP-14)催化结构域为靶标,通过噬菌体随机十二肽库筛选和分子模拟、细胞免疫荧光、金属离子亲和层析以及体外细胞作用测定等技术,进行了双靶向MMP-14和金属离子小分子结合多肽的筛选与研究.经4轮筛选,噬菌体得到有效富集并获得13条不同的多肽序列.序列分析显示,可能的一致序列有:AHQLH、HHXH、EI/LPLL/I.分子模拟与对接进一步确认一致序列AHQLH、HHTH、LPLL与MMP-14催化结构域的氨基酸120-125区域良好分子对接并具有一定的专一性,多条MMP-14结合肽不仅靶向MMP-14,同时结合金属离子.细胞生物学研究确认,所测定的结合肽噬菌体对MMP-14诱导表达的MG63细胞具有良好的结合作用,揭示结合肽对MMP-14的靶向结合特性,并且合成的AHQLH、LPLL一致序列多肽对MG63细胞活力具有一定的抑制能力.这些新的和具有一定MMP-14专一性的一致序列可望用于靶向MMP-14抗肿瘤药物的研发和利用.election and study of lead binding peptides dual targeting matrix metalloproteinase-14（MMP-14） and metal ions were carried out by phage random dodecapepetide library selection against catalytic domain of MMP-14,molecular simulation,cell immunofluorescence,metal ion affinity chromatography and in vitro cell assay.Phages were efficiently enriched after four rounds of selection and totally 13 different peptide sequences were obtained.The possible peptide binding consensus sequences were found as AHQLH,HHXH and EI/LPLL/I.Molecular simulation and docking further confirmed that the consensus sequences AHQLH,HHTH and LPLL can dock well to the targeted catalytic domains of MMP-14 at the sites of amino acids 120～125 and are relatively specific for MMP-14.Meanwhile,it wasfound that several MMP-14 binding peptides were not only targeted to MMP14 but also metal ions.Cellbiological study showed that several assayed monoclonal MMP-14 phages containing binding peptidescould bind well to the MG-63 cells which were induced for expression of MMP-14 and this againconfirmed the targeting binding of selected peptides to MMP-14.In addition,the selected and chemically synthesized consensus peptides had potential in inhibiting the viability of MG-63 cells in vitro.Theconsensus sequences were novel and relatively specific to MMP14,and they may be used in both the basic research and lead peptides for MMP-14 based cancer therapeutic drug development.

关 键 词：基质金属蛋白酶-14(MMP-14) 噬菌体随机十二肽库 一致序列 对接 分子模拟 

分 类 号：Q5[生物学—生物化学] Q7