• Protrudin-mediated ER-endosome contact sites promote MT1-MMP exocytosis and cell invasion.

Protrudin-mediated ER-endosome contact sites promote MT1-MMP exocytosis and cell invasion.

The Journal of cell biology (2020-06-02)
Nina Marie Pedersen, Eva Maria Wenzel, Ling Wang, Sandra Antoine, Philippe Chavrier, Harald Stenmark, Camilla Raiborg

Cancer cells break tissue barriers by use of small actin-rich membrane protrusions called invadopodia. Complete invadopodia maturation depends on protrusion outgrowth and the targeted delivery of the matrix metalloproteinase MT1-MMP via endosomal transport by mechanisms that are not known. Here, we show that the ER protein Protrudin orchestrates invadopodia maturation and function. Protrudin formed contact sites with MT1-MMP-positive endosomes that contained the RAB7-binding Kinesin-1 adaptor FYCO1, and depletion of RAB7, FYCO1, or Protrudin inhibited MT1-MMP-dependent extracellular matrix degradation and cancer cell invasion by preventing anterograde translocation and exocytosis of MT1-MMP. Moreover, when endosome translocation or exocytosis was inhibited by depletion of Protrudin or Synaptotagmin VII, respectively, invadopodia were unable to expand and elongate. Conversely, when Protrudin was overexpressed, noncancerous cells developed prominent invadopodia-like protrusions and showed increased matrix degradation and invasion. Thus, Protrudin-mediated ER-endosome contact sites promote cell invasion by facilitating translocation of MT1-MMP-laden endosomes to the plasma membrane, enabling both invadopodia outgrowth and MT1-MMP exocytosis.

Product Number
Product Description

Hepatocyte Growth Factor human, HGF, recombinant, expressed in HEK 293 cells, HumanKine®, suitable for cell culture
Anti-SH3PXD2A antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution
Corning® Costar® Ultra-Low Attachment Multiple Well Plate, size 96 well, round bottom clear, pkg of (individually wrapped), pkg of (24/case), sterile, lid
PIPES dipotassium salt, ≥99% (titration)
Saponin from quillaja bark, Sapogenin content ≥10 %
Triton X-100, laboratory grade