The transmembrane domain is essential for the microtubular trafficking of membrane type-1 matrix metalloproteinase (MT1-MMP).
Membrane type-1 matrix metalloproteinase (MT1-MMP) degrades the extracellular matrix, initiates the activation pathway of soluble MMPs and regulates the functionality of cell adhesion signaling receptors, thus playing an important role in many cell functions. Intracellular transport mechanisms, currently incompletely understood, regulate the presentation of MT1-MMP at the cell surface. We have focused our efforts on identifying these mechanisms. To understand the transport of MT1-MMP across the cell, we used substitution and deletion mutants, the trafficking of which was examined using antibody uptake and Chariot delivery experiments. Our experiments have demonstrated that the microtubulin cytoskeleton and the centrosomes (the microtubulin cytoskeleton-organizing centers) are essential for the trafficking and the internalization of MT1-MMP. We determined that after reaching the plasma membrane, MT1-MMP is internalized in the Rab-4-positive recycling endosomes and the Rab-11-positive pericentrosomal recycling endosomes. The microtubular trafficking causes the protease to accumulate in the pericentrosomal region of the cell. We believe that the presence of the transmembrane domain is required for the microtubular vesicular trafficking of MT1-MMP because the soluble mutants are not presented at the cell surface and they are not delivered to the centrosomes. The observed transport mechanisms provide a vehicle for the intracellular targets and, accordingly, for an intracellular cleavage function of MT1-MMP in malignant cells, which routinely overexpress this protease.
Remacle
Albert G
AG
The Burnham Institute, La Jolla, CA 92037, USA.
Rozanov
Dmitri V
DV
Baciu
Peter C
PC
Chekanov
Alexei V
AV
Golubkov
Vladislav S
VS
Strongin
Alex Y
AY
D000818:Animals; D001666:Binding Sites, Antibody; D016466:CHO Cells; D002460:Cell Line; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000201:enzymology*; D018385:Centrosome / Q000201:enzymology / Q000378:metabolism; D006224:Cricetinae; D003599:Cytoskeleton / Q000502:physiology; D004285:Dogs; D006801:Humans; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D008870:Microtubules / Q000201:enzymology* / Q000378:metabolism; D017434:Protein Structure, Tertiary / Q000502:physiology; D021381:Protein Transport / Q000235:genetics / Q000502:physiology; D017384:Sequence Deletion; D022161:Transport Vesicles / Q000201:enzymology / Q000235:genetics; D014404:Tubulin / Q000502:physiology
D014404:Tubulin; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases