Cytoplasmic tail–dependent internalization of membrane-type 1 matrix metalloproteinase is important for its invasion-promoting activity

Abstract

Membrane-type 1 matrix metalloproteinase (MT1-MMP) is an integral membrane proteinase that degrades the pericellular extracellular matrix (ECM) and is expressed in many migratory cells, including invasive cancer cells. MT1-MMP has been shown to localize at the migration edge and to promote cell migration; however, it is not clear how the enzyme is regulated during the migration process. Here, we report that MT1-MMP is internalized from the surface and that this event depends on the sequence of its cytoplasmic tail. Di-leucine (Leu571–572 and Leu578–579) and tyrosine573 residues are important for the internalization, and the μ2 subunit of adaptor protein 2, a component of clathrin-coated pits for membrane protein internalization, was found to bind to the LLY573 sequence. MT1-MMP was internalized predominantly at the adherent edge and was found to colocalize with clathrin-coated vesicles. The mutations that disturb internalization caused accumulation of the enzyme at the adherent edge, though the net proteolytic activity was not affected much. Interestingly, whereas expression of MT1-MMP enhances cell migration and invasion, the internalization-defective mutants failed to promote either activity. These data indicate that dynamic turnover of MT1-MMP at the migration edge by internalization is important for proper enzyme function during cell migration and invasion.

Hic-5 mediates endothelial sprout initiation by regulating a key surface metalloproteinase

“…For this reason, we performed surface biotinylation assays, which demonstrated decreased surface expression of MT1-MMP that correlated with decreased invasion responses in the absence of Hic-5. Thus, Hic-5 silencing did not alter overall MT1-MMP activity levels, supporting that surface localization of MT1-MMP, rather than overall activity, is a key indicator of invasive behavior (Hotary et al, 2000;Yana and Weiss, 2000;Uekita et al, 2001;Egeblad and Werb, 2002;Li et al, 2008;Kwak et al, 2012). The observation that Hic-5 depletion decreased S1P-mediated MT1-MMP plasma membrane translocation in surface biotinylation assays, suggests that Hic-5 can function as a molecular adaptor to transport activated MT1-MMP to the plasma membrane for successful sprouting responses.…”

“…In addition to Hic-5, invadopodia also contain membrane type-1 matrix metalloproteinase (MT1-MMP, also known as MMP14) (Egeblad and Werb, 2002). MT1-MMP is a member of the MMP family that contains a transmembrane domain, which anchors it to the cell surface and facilitates the proximal extracellular matrix (ECM) breakdown needed for invasion into 3D matrices (Hotary et al, 2000;Yana and Weiss, 2000;Uekita et al, 2001;Egeblad and Werb, 2002;Li et al, 2008). Although multiple classes of MMPs have been implicated in angiogenic events, MT1-MMP is essential for endothelial invasion through a 3D ECM.…”

Metalloproteinases and their inhibitors in tumor angiogenesis

“…[12][13][14][15] Intracellular proteins may regulate the localization and activity of MT1-MMP by interacting with its cytoplasmic tail, potentially via dileucine and tyrosine residues within this domain. [16][17][18] MT-MMP-4 and -6 are GPI-anchored. 19,20 …”

“…12-15 Intracellular proteins may regulate the localization and activity of MT1-MMP by interacting with its cytoplasmic tail, potentially via dileucine and tyrosine residues within this domain. [16][17][18]20 Endogenous inhibitors of metalloproteinasesThe activity of MMPs can be regulated by endogenous inhibitors, which include the TIMPs, 21 a 2 -macroglobulin 22 and the membrane-anchored glycoprotein RECK (for ''reversioninducing cysteine-rich protein with kazal motifs''). 23 There are 4 vertebrate TIMPs.…”

Membrane-Type 1 Matrix Metalloproteinase Stimulates Cell Migration through Epidermal Growth Factor Receptor Transactivation

“…The overexpression of MT1-MMP enables tumor cell growth in otherwise growthrestrictive, three-dimensional extracellular matrices (7) and promotes cell migration of a number of cancer (8)(9)(10) and endothelial cell lines (11)(12)(13). The short cytoplasmic sequence of MT1-MMP also contributes to the induction of cell migration by the enzyme (9, 10, 13) possibly through activation of the extracellular signal-regulated protein kinase (ERK) cascade (14), endocytosis and trafficking of the enzyme (15,16), and interaction with tyrosine phosphorylated caveolin-1 (17). Additional insights into the mechanisms by which MT1-MMP induces cell migration may come from our observation that MT1-MMP cooperates with the plateletderived bioactive lipid sphingosine 1-phosphate (S1P) to stimulate endothelial cell migration and morphogenic differentiation into capillary-like structures in vitro (13).…”