Molecular Biology Department, Universidad Autónoma de Madrid (UAM), 28049 Madrid, Spain.
Severo Ochoa Molecular Biology Center (CBM-SO), Instituto de Investigación Sanitaria Princesa (IIS-IP), 28049 Madrid, Spain.
Cells. 2020 Feb 3;9(2):348. doi: 10.3390/cells9020348.
Membrane-bound proteases play a key role in biology by degrading matrix proteins or shedding adhesion receptors. MT1-MMP metalloproteinase is critical during cancer invasion, angiogenesis, and development. MT1-MMP activity is strictly regulated by internalization, recycling, autoprocessing but also through its incorporation into tetraspanin-enriched microdomains (TEMs), into invadopodia, or by its secretion on extracellular vesicles (EVs). We identified a juxtamembrane positively charged cluster responsible for the interaction of MT1-MMP with ERM (ezrin/radixin/moesin) cytoskeletal connectors in breast carcinoma cells. Linkage to ERMs regulates MT1-MMP subcellular distribution and internalization, but not its incorporation into extracellular vesicles. MT1-MMP association to ERMs and insertion into TEMs are independent phenomena, so that mutation of the ERM-binding motif in the cytoplasmic region of MT1-MMP does not preclude its association with the tetraspanin CD151, but impairs the accumulation and coalescence of CD151/MT1-MMP complexes at actin-rich structures. Conversely, gene deletion of CD151 does not impact on MT1-MMP colocalization with ERM molecules. At the plasma membrane MT1-MMP autoprocessing is severely dependent on ERM association and seems to be the dominant regulator of the enzyme collagenolytic activity. This newly characterized MT1-MMP/ERM association can thus be of relevance for tumor cell invasion.
膜结合蛋白酶通过降解基质蛋白或脱落黏附受体在生物学中发挥关键作用。MT1-MMP 金属蛋白酶在癌症侵袭、血管生成和发育过程中至关重要。MT1-MMP 的活性受到严格调控,包括内化、循环、自身加工,但也包括其整合到富含四跨膜蛋白的微区(TEM)中、进入入侵伪足中,或通过其分泌到细胞外囊泡(EV)中。我们鉴定了一个位于跨膜区附近的正电荷簇,该簇负责 MT1-MMP 与乳腺癌细胞中 ERM(ezrin/radixin/moesin)细胞骨架连接蛋白的相互作用。与 ERMs 的连接调节 MT1-MMP 的亚细胞分布和内化,但不调节其整合到细胞外囊泡中。MT1-MMP 与 ERMs 的关联和插入 TEM 是独立的现象,因此 MT1-MMP 细胞质区域中 ERM 结合基序的突变并不排除其与四跨膜蛋白 CD151 的关联,但会损害 CD151/MT1-MMP 复合物在富含肌动蛋白的结构中的积累和融合。相反,CD151 的基因缺失并不影响 MT1-MMP 与 ERM 分子的共定位。在质膜上,MT1-MMP 的自身加工严重依赖于 ERM 的关联,并且似乎是该酶胶原酶活性的主要调节剂。因此,新表征的 MT1-MMP/ERM 关联可能与肿瘤细胞侵袭有关。