Matrix metalloproteinases (MMPs) are evolutionarily conserved extracellular matrix proteinases. Genetic analysis of the Drosophila MMPs, Mmp1 and Mmp2, in vivo reveal that they play vital roles in tissue remodeling. Although the catalytic domain (CD) undertakes most MMP functions, few studies have sought to demonstrate the biochemical properties of the CDs of fly MMPs. Here, we identified the overexpression, purification, and refolding of the CDs of Drosophila Mmp1 and Mmp2 for biochemical studies. Zymography assays and substrate degradation analysis showed that both Mmp1-CD and Mmp2-CD were able to digest casein, gelatin, fibronectin, collagen (types I, IV, and V), while Mmp2-CD showed much higher degradation activity compared with Mmp1-CD. Moreover, human collagen III could be degraded by Mmp1-CD but not Mmp2-CD and rat collagen I and laminin could be degraded by Mmp2-CD but not Mmp1-CD, suggesting that Drosophila Mmp1 and Mmp2 might have overlapping yet distinct substrate specificity. Using synthetic fluorescent substrates, we further demonstrated that the enzymatic activity of Mmp1-CD and Mmp2-CD could be inhibited by human tissue inhibitors of metalloproteinases (TIMPs). These results reveal the context of the cooperative yet distinct roles of Mmp1 and Mmp2 in tissue remodeling.

基质金属蛋白酶（MMPs）是进化上保守的细胞外基质蛋白酶。果蝇MMPs Mmp1和Mmp2在体内的遗传分析表明，它们在组织重塑中起着至关重要的作用。尽管催化域（CD）承担大多数MMP功能，但很少有研究试图证明苍蝇MMP的CD的生化特性。在这里，我们确定了果蝇CD的过表达，纯化和重折叠Mmp1和Mmp2用于生化研究。阻抗谱分析和底物降解分析表明，Mmp1-CD和Mmp2-CD都能消化酪蛋白，明胶，纤连蛋白，胶原蛋白（I，IV和V型），而Mmp2-CD的降解活性比Mmp1-CD高得多。光盘。此外，人胶原蛋白III可以被Mmp1-CD降解，但不能被Mmp2-CD降解，大鼠胶原蛋白I和层粘连蛋白可以被Mmp2-CD降解，而不能被Mmp1-CD降解，这表明果蝇Mmp1和Mmp2可能具有重叠但截然不同的底物特异性。使用合成的荧光底物，我们进一步证明了Mmp1-CD和Mmp2-CD的酶活性可以被人类金属蛋白酶抑制剂（TIMPs）抑制。这些结果揭示了在组织重塑中Mmp1和Mmp2的协同作用但又有独特作用的背景。