Dauvé Jonathan, Belloy Nicolas, Rivet Romain, Etique Nicolas, Nizet Pierre, Pietraszek-Gremplewicz Katarzyna, Karamanou Konstantina, Dauchez Manuel, Ramont Laurent, Brézillon Stéphane, Baud Stéphanie
P3M, Multi-Scale Molecular Modeling Platform, Université de Reims Champagne Ardenne, 51097 Reims, France.
CNRS UMR 7369, Matrice Extracellulaire et Dynamique Cellulaire (MEDyC), Université de Reims Champagne Ardenne, 51095 Reims, France.
Cancers (Basel). 2021 Sep 30;13(19):4930. doi: 10.3390/cancers13194930.
Lumican, a small leucine-rich proteoglycan (SLRP) of the extracellular matrix (ECM), displays anti-tumor properties through its direct interaction with MMP-14. Lumican-derived peptides, such as lumcorin (17 amino acids) or L9M (10 amino acids), are able to inhibit the proteolytic activity of MMP-14 and melanoma progression. This work aimed to visualize the interactions of lumican-derived peptides and MMP-14. Molecular modeling was used to characterize the interactions between lumican-derived peptides, such as lumcorin, L9M, and cyclic L9M (L9Mc, 12 amino acids), and MMP-14. The interaction of L9Mc with MMP-14 was preferential with the MT-Loop domain while lumcorin interacted more with the catalytic site. Key residues in the MMP-14 amino acid sequence were highlighted for the interaction between the inhibitory SLRP-derived peptides and MMP-14. In order to validate the in silico data, MMP-14 activity and migration assays were performed using murine B16F1 and human HT-144 melanoma cells. In contrast to the HT-144 melanoma cell line, L9Mc significantly inhibited the migration of B16F1 cells and the activity of MMP-14 but with less efficacy than lumican and lumcorin. L9Mc significantly inhibited the proliferation of B16F1 but not of HT-144 cells in vitro and primary melanoma tumor growth in vivo. Thus, the site of interaction between the domains of MMP-14 and lumcorin or L9Mc were different, which might explain the differences in the inhibitory effect of MMP-14 activity. Altogether, the biological assays validated the prediction of the in silico study. Possible and feasible improvements include molecular dynamics results.
纤连蛋白是细胞外基质（ECM）中的一种富含亮氨酸的小分子蛋白聚糖（SLRP），通过与基质金属蛋白酶-14（MMP-14）直接相互作用展现出抗肿瘤特性。纤连蛋白衍生肽，如纤连蛋白（17个氨基酸）或L9M（10个氨基酸），能够抑制MMP-14的蛋白水解活性以及黑色素瘤进展。本研究旨在可视化纤连蛋白衍生肽与MMP-14的相互作用。采用分子建模来表征纤连蛋白衍生肽，如纤连蛋白、L9M和环L9M（L9Mc，12个氨基酸）与MMP-14之间的相互作用。L9Mc与MMP-14的相互作用在MT-环结构域处更具优势，而纤连蛋白与催化位点的相互作用更多。突出了MMP-14氨基酸序列中的关键残基，以用于抑制性SLRP衍生肽与MMP-14之间的相互作用。为了验证计算机模拟数据，使用小鼠B16F1和人HT-144黑色素瘤细胞进行了MMP-14活性和迁移试验。与HT-144黑色素瘤细胞系不同，L9Mc显著抑制了B16F1细胞的迁移和MMP-14的活性，但效果不如纤连蛋白和纤连蛋白。L9Mc在体外显著抑制了B16F1细胞的增殖，但对HT-144细胞无此作用，且在体内抑制了原发性黑色素瘤肿瘤的生长。因此，MMP-14结构域与纤连蛋白或L9Mc之间的相互作用位点不同，这可能解释了对MMP-14活性抑制作用的差异。总之，生物学试验验证了计算机模拟研究的预测。可能且可行的改进包括分子动力学结果。
