Shedding of the 67-kD laminin receptor by human cancer cells.
The 67-kD laminin receptor (67LR) is a cell membrane-associated molecule exhibiting high affinity for the basement membrane glycoprotein, laminin. While export of the 67LR toward the extracellular matrix has been recently suggested by electron microscopy studies, there is to date no evidence of shedding of the 67LR from cells. Using two monoclonal antibodies directed against the 67LR, we developed a double-determinant radioimmunoassay that demonstrates that the 67LR is released from cancer cells into the culture medium. The shed molecule exhibited the same apparent molecular weight as that of the membrane-associated 67LR, suggesting that no proteolytic cleavage is involved in the process. Furthermore, we demonstrate that the 67LR is not anchored to the membrane through a glycolsyl-phosphatidylinositol bridge. However, the observation that lactose increased the release of 67LR suggests that a lectin-type interaction is involved in the cell membrane association of this laminin binding protein and the cell surface. Interestingly, the released 67LR recovered after HPLC gel filtration was found free as well as associated to high molecular weight complexes. The free 67LR retained its ability to bind to the cell surface. Our study is the first demonstration that the 67LR is effectively shed by cancer cells. The released free 67LR could play an important role in modulating interactions between cancer cells and laminin during tumor invasion and metastasis.
Karpatová
M
M
Division of Experimental Oncology E, Istituto Nazionale per lo Studio e la Cura dei Tumori, Milan, Italy.
Tagliabue
E
E
Castronovo
V
V
Magnifico
A
A
Ardini
E
E
Morelli
D
D
Belotti
D
D
Colnaghi
M I
MI
Ménard
S
S
D000911:Antibodies, Monoclonal; D002462:Cell Membrane / Q000378:metabolism; D003470:Culture Media; D005109:Extracellular Matrix / Q000378:metabolism*; D006801:Humans; D008970:Molecular Weight; D011863:Radioimmunoassay / Q000379:methods; D017474:Receptors, Laminin / Q000378:metabolism*; D012995:Solubility; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D003470:Culture Media; D017474:Receptors, Laminin