靶向基质金属蛋白酶14类血红素结构域特异活性位点的确定及反义肽虚拟筛选

目的:确定基质金属蛋白酶14(MMP-14)类血红素结构域(HPX14)特异活性位点,虚拟筛选获得能与HPX14特异结合的短肽。方法:用metaPocket预测HPX14的活性口袋,多序列比对分析活性口袋的氨基酸残基特异性;基于M-I和R-B理论设计以特异活性位点为正义肽的反义肽库,并进行虚拟筛研究及结合特异性确定。结果:生物信息学分析确定了KGRGLTD为HPX14的特异活性位点,并构建了其1036条反义肽;通过2轮虚拟筛选,获得10条得分居前的与HPX14结合较好的反义肽,它们与HPX14具有较高的亲和力,并可能影响MMP-14同源二聚体的形成和MMP-14活性的抑制;VSETAPF、IGEPPPF是对接打分最好的2条短肽,与HPX14的结合具有特异性。结论:KGRGLTD是HPX14的特异活性位点,虚拟筛选得到的VSETAPF、IGEPPPF等反义肽与HPX14具有较高的亲和力,这为基于MMP-14的HPX结构域的靶向小分子多肽先导药物的发现提供了重要的前期工作基础与新思路。

Identification of the Specific Active Site of MMP-14 Hemopexin-Like Domain and the Targeted Virtual Screening of Antisense Peptides

Objective： To find the specific active site of matrix metalloproteinase 14（MMP-14） hemopexin-like domain（HPX14） and obtain HPX14 specific binding peptides by virtual screening. Methods： The active pockets of HPX14 were predicted by metaPocket and the specificity of residues in the active pocket was analyzed by multiple sequence alignment. Based on M-I and R-B theory, an antisense peptide libraries against the active site（as the sense peptide） was designed, then virtual screening and binding specificity study were performed. Results： KGRGLTD as the specific active site of HPX14 was identified by bioinformaties analysis and 1036 antisense peptides against it was designed. By two rounds of virtual screening, the top 10 peptides ranked by the predicted affinity for HPX14 were obtained. Those peptides have comparatively higher affinities to HPX14 and may affect the homodimerization of MMP-14 and finally the inhibition of MMP-14 activity. VSETAPF, IGEPPPF were the top 2 highest scored peptides and can specifically bind to HPX14. Conclusion： KGRGLTD was the specific active site of the HPX14. VSETAPF, IGEPPPF and other antisense peptides that obtained by virtual screening have high affinity to HPX14. The work provides an important base as well as new ideas and approach for the discovery and study of specific lead peptides targeting HPX14.