PDF(Pubmed)
Abstract：
Angiogenesis plays an essential role in various normal physiological processes, such as embryogenesis, tissue repair, and skin regeneration. Visfatin is a 52 kDa adipokine secreted by various tissues including adipocytes. It stimulates the expression of vascular endothelial growth factor (VEGF) and promotes angiogenesis. However, there are several issues in developing full-length visfatin as a therapeutic drug due to its high molecular weight. Therefore, the purpose of this study was to develop peptides, based on the active site of visfatin, with similar or superior angiogenic activity using computer simulation techniques.Initially, the active site domain (residues 181∼390) of visfatin was first truncated into small peptides using the overlapping technique. Subsequently, the 114 truncated small peptides were then subjected to molecular docking analysis using two docking programs (HADDOCK and GalaxyPepDock) to generate small peptides with the highest affinity for visfatin. Furthermore, molecular dynamics simulations (MD) were conducted to investigate the stability of the protein-ligand complexes by computing root mean square deviation (RSMD) and root mean square fluctuation(RMSF) plots for the visfatin-peptide complexes. Finally, peptides with the highest affinity were examined for angiogenic activities, such as cell migration, invasion, and tubule formation in human umbilical vein endothelial cells (HUVECs). Through the docking analysis of the 114 truncated peptides, we screened nine peptides with a high affinity for visfatin. Of these, we discovered two peptides (peptide-1: LEYKLHDFGY and peptide-2: EYKLHDFGYRGV) with the highest affinity for visfatin. In an in vitrostudy, these two peptides showed superior angiogenic activity compared to visfatin itself and stimulated mRNA expressions of visfatin and VEGF-A. These results show that the peptides generated by the protein-peptide docking simulation have a more efficient angiogenic activity than the original visfatin.
摘要：
血管生成在各种正常生理过程中起着至关重要的作用,比如胚胎发生，组织修复,和皮肤再生。内脂素是由包括脂肪细胞的各种组织分泌的52kDa脂肪因子。它刺激血管内皮生长因子（VEGF）的表达并促进血管生成。然而,由于其分子量高，在开发全长visfatin作为治疗药物方面存在几个问题。因此,这项研究的目的是开发肽，基于visfatin的活性位点，使用计算机模拟技术具有相似或优越的血管生成活性。最初，首先使用重叠技术将内脂素的活性位点结构域（残基181~390）截短为小肽。随后，然后使用两个对接程序(HADDOCK和GalaxyPepDock)对114个截短的小肽进行分子对接分析,以产生对内脂素具有最高亲和力的小肽.此外，分子动力学模拟（MD）通过计算内脂素-肽复合物的均方根偏差（RSMD）和均方根波动（RMSF）图来研究蛋白质-配体复合物的稳定性。最后,检查具有最高亲和力的肽的血管生成活性，如细胞迁移，入侵,和人脐静脉内皮细胞（HUVECs）的小管形成。通过对114个截短肽的对接分析，我们筛选了9种对内脂素具有高亲和力的肽。其中,我们发现了对内脂素具有最高亲和力的两种肽（肽-1：LEYKLHDFGY和肽-2：EYKLHDFGYRGV）。在体外研究中，与内脂素本身相比，这两种肽显示出优异的血管生成活性，并刺激内脂素和VEGF-A的mRNA表达。这些结果表明,通过蛋白质-肽对接模拟产生的肽具有比原始内脂素更有效的血管生成活性。
