Deciphering MMP9's dual role in regulating SOD3 through protein-protein interactions.
Deciphering MMP9's dual role in regulating SOD3 through protein-protein interactions.
复制标题
解读 MMP9 通过蛋白质-蛋白质相互作用调节 SOD3 的双重作用。
DOI：
10.1139/cjpp-2023-0256
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发表时间：
2024
影响因子：
2.1
通讯作者：
Mishra,ParasK
中科院分区：
文献类型：
--
作者：
Gawargi,FlobaterI;Mishra,ParasK
尽管基质金属蛋白酶9（MMP 9）的胶原酶活性已得到充分证实，但其非酶功能仍不太清楚。细胞内超氧化物歧化酶-1（SOD 1）和MMP 9之间的相互作用是已知的，SOD 1抑制MMP 9。然而，MMP 9作为一种分泌性蛋白，影响细胞外抗氧化剂超氧化物歧化酶-3（SOD-3）的机制尚不清楚。为了探索MMP 9对SOD 3的调节作用，我们采用人胚肾-293细胞，用MMP 9过表达和催化位点突变质粒转染它们。此外，用MMP 9激活剂和抑制剂处理MMP 9过表达细胞。对细胞裂解物和培养基的分析提供了对MMP 9的细胞内和细胞外调节作用的见解。利用计算机模拟分析和实验方法如近端连接测定和免疫共沉淀来描绘MMP 9和SOD 3之间的蛋白质-蛋白质相互作用。我们的研究结果表明，活化的MMP 9增强了SOD 3水平，这是一种不受MMP 9抑制剂阻碍的调节。有趣的是，无催化活性的MMP 9似乎降低了SOD 3水平，这可能是由于MMP 9与SOD 3结合，导致其蛋白水解降解。这种MMP 9对SOD 3的影响在细胞内和细胞外环境中是一致的，表明MMP 9-SOD 3相互作用在这些结构域中是平行的。最终，这项研究揭示了MMP 9和SOD 3之间的一种新的相互作用，突出了催化失活的MMP 9在降低SOD 3水平方面的独特调节作用，与活性MMP 9的通常上调形成对比。
Although the collagenase enzyme activity of matrix metalloproteinase-9 (MMP9) is well-documented, its non-enzymatic functions remain less understood. The interaction between intracellular superoxide dismutase-1 (SOD1) and MMP9 is known, with SOD1 suppressing MMP9. However, the mechanism by which MMP9, a secretory protein, influences the extracellular antioxidant superoxide dismutase-3 (SOD3) is not yet clear. To explore MMP9's regulatory impact on SOD3, we employed human embryonic kidney-293 cells, transfecting them with MMP9 overexpresssion and catalytic-site mutant plasmids. Additionally, MMP9 overexpressing cells were treated with an MMP9 activator and inhibitor. Analyses of both cell lysates and culture medium provided insights into MMP9's intracellular and extracellular regulatory roles. In-silico analysis and experimental approaches like proximal ligation assay and co-immunoprecipitation were utilized to delineate the protein-protein interactions between MMP9 and SOD3. Our findings indicate that activated MMP9 enhances SOD3 levels, a regulation not hindered by MMP9 inhibitors. Intriguingly, catalytically inactive MMP9 appeared to reduce SOD3 levels, likely due to MMP9's binding with SOD3, leading to their proteolytic degradation. This MMP9 influence on SOD3 was consistent in both intracellular and extracellular environments, suggesting a parallel in MMP9-SOD3 interactions across these domains. Ultimately, this study unveils a novel interaction between MMP9 and SOD3, highlighting the unique regulatory role of catalytically inactive MMP9 in diminishing SOD3 levels, contrasting its usual upregulation by active MMP9.