Identification of laminin binding proteins in cell membranes of a human colon adenocarcinoma cell line.
The invasion of malignant cells through the basement membrane is a critical step in local infiltration and metastasis. Adhesion and invasion of malignant cells may be modulated by their receptor mediated binding to the basement membrane glycoprotein laminin. We studied the specific adhesion of human colon adenocarcinoma derived HT 29 cells to laminin and its proteolytic fragments. The major cell adhesion domain of laminin was localised in the central part of the cross shaped molecule. Immunoblotting experiments on separated HT 29 cell membranes using specific antibodies or radiolabelled laminin fragments revealed two major laminin-binding cell surface components with Mr of 67,000 and 69,000 D similar to the putative laminin receptor described for other tissues. Using a nitrocellulose filter disk assay, the specific interaction between cell surface binding proteins and proteolytic fragments originating from the central core of the laminin molecule could be further corroborated. In contrast, interaction of HT 29 cell membranes with the pentapeptide YIGSR (tyr-ile-gly-ser-arg), a sequence domain of the B1-chain of the laminin molecule, thought to be responsible for cell adhesion, was significantly weaker.
Stallmach
A
A
Freie Universität, Berlin, Federal Republic of Germany.
Schuppan
D
D
Dax
J
J
Hanski
C
C
Riecken
E O
EO
D000230:Adenocarcinoma / Q000032:analysis*; D002460:Cell Line; D002462:Cell Membrane / Q000032:analysis; D003110:Colonic Neoplasms / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D015151:Immunoblotting; D007797:Laminin; D008970:Molecular Weight; D011946:Receptors, Antigen / Q000032:analysis*; D011971:Receptors, Immunologic / Q000032:analysis*; D017474:Receptors, Laminin; D014407:Tumor Cells, Cultured
D007797:Laminin; D011946:Receptors, Antigen; D011971:Receptors, Immunologic; D017474:Receptors, Laminin