Zeng Li, Tan Bowen, Yang Yalan, Qiu Jinyi, Xiong Lili, Mao Canquan
Sheng Wu Gong Cheng Xue Bao. 2015 Feb;31(2):269-80.
Membrane type-1 matrix metalloproteinase (MT1-MMP or MMP14) plays the pivotal role in tumor development and metastasis, so it is a promising drug target in malignancy. To acquire MT1-MMP specific binding peptides, we first analyzed MMPs sequences to find the divergent and specific sequence of MT1-MMP by bioinformatics approach, then set the specific sequence as the sense peptide target and designed antisense peptide library. Finally, by means of molecular docking, molecular dynamics simulation and in vitro cell assays, we screened the antisense peptide library against MT1-MMP and further studied the obtained specific peptides. Here, we identified the divergent and specific sequence of AYIREGHE (Named MT1-loop) located in MT1-MMP loop by multiple sequence alignment and established the antisense peptides library with capacity of 1 536 sequences. After two rounds of virtual screening, we obtained five antisense peptides with Rerankscores in the top for further screening. They all interacted with MT1-MMP, and docked well at the active site composed of MT1-loop sequence. Analysis of the affinities of these five antisense peptides to other MMPs (MMP1-3, MMP7-13, MMP14 HPX, MMP16) revealed that the peptide FVTFPYIR was more specific to MT1-MMP. Molecular dynamics simulation showed that the peptide FVTFPYIR might affect the stability of MT1-MMP and thus have effects on its activities. Meanwhile, the peptide FVTFPYIR could specifically inhibit the growth of MG63 and MDA-MB-231 tumor cells both of which expressed MT1-MMP. The work provides a new insight and way for the development of antitumor lead peptides targeting MT1-MMP.
膜型-1基质金属蛋白酶（MT1-MMP或MMP14）在肿瘤发生和转移中起关键作用，因此它是恶性肿瘤中一个有前景的药物靶点。为了获得MT1-MMP特异性结合肽，我们首先通过生物信息学方法分析基质金属蛋白酶序列以找到MT1-MMP的差异和特异性序列，然后将该特异性序列设定为有义肽靶点并设计反义肽库。最后，通过分子对接、分子动力学模拟和体外细胞实验，我们针对MT1-MMP筛选了反义肽库并进一步研究获得的特异性肽。在此，我们通过多序列比对确定了位于MT1-MMP环中的AYIREGHE差异和特异性序列（命名为MT1-环），并建立了具有1536个序列容量的反义肽库。经过两轮虚拟筛选，我们获得了五个重排分数排名靠前的反义肽用于进一步筛选。它们均与MT1-MMP相互作用，并在由MT1-环序列组成的活性位点处对接良好。对这五个反义肽与其他基质金属蛋白酶（MMP1-3、MMP7-13、MMP14 HPX、MMP16）亲和力的分析表明，肽FVTFPYIR对MT1-MMP更具特异性。分子动力学模拟表明，肽FVTFPYIR可能影响MT1-MMP的稳定性，从而对其活性产生影响。同时，肽FVTFPYIR可特异性抑制表达MT1-MMP的MG63和MDA-MB-231肿瘤细胞的生长。这项工作为开发靶向MT1-MMP的抗肿瘤先导肽提供了新的见解和途径。
