Extracellular matrix protein 1 inhibits the activity of matrix metalloproteinase 9 through high-affinity protein/protein interactions
Extracellular matrix protein 1 inhibits the activity of matrix metalloproteinase 9 through high-affinity protein/protein interactions
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DOI：
10.1111/j.0906-6705.2006.00409.x
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发表时间：
2006-04-01
影响因子：
3.6
通讯作者：
Uitto, J
中科院分区：
文献类型：
--
作者：
Fujimoto, N;Terlizzi, J;Uitto, J
细胞外基质蛋白1（ECM 1）是一种约85 kDa的糖蛋白，具有广泛的组织分布，在类脂质蛋白沉积症（LP）中存在突变，LP是一种遗传性疾病，其特征为基底膜重复和真皮透明化，与神经系统疾病相关。从ECM 1突变到LP表型的机制尚不清楚。本研究利用酵母双杂交技术对人胎盘组织中ECM 1蛋白相互作用进行了研究，筛选出9个相互作用蛋白，其中包括基质金属蛋白酶9（MMP 9）。通过分离克隆的β-半乳糖苷酶测定和将ECM 1中的相互作用片段缩小至C末端串联重复序列2（氨基酸236-361）的免疫共沉淀证实了相互作用。该肽段还在基于明胶的ELISA测定中抑制MMP 9活性。我们认为ECM 1介导的MMP 9蛋白水解活性的降低可能与LP的发病机制有关。
Extracellular matrix protein 1 (ECM1), an approximately 85-kDa glycoprotein with broad tissue distribution, harbors mutations in lipoid proteinosis (LP), a heritable disease characterized by reduplication of basement membranes and hyalinization of dermis, associated with neurologic disorders. The mechanisms leading from ECM1 mutations to LP phenotype are unknown. In this study, we explored ECM1 protein-protein interactions utilizing yeast two-hybrid genetic screen of human placental library, which identified nine interacting proteins, including matrix metalloproteinase 9 (MMP9). The interactions were confirmed by beta-galactosidase assay with isolated clones and by co-immunoprecipitation which narrowed the interacting segment in ECM1 to the C-terminal tandem repeat 2 (amino acids 236-361). This peptide segment also inhibited MMP9 activity in a gelatin-based ELISA assay. We propose that ECM1-mediated reduction in MMP9 proteolytic activity may have relevance to pathogenesis of LP.