Force required to break alpha5beta1 integrin-fibronectin bonds in intact adherent cells is sensitive to integrin activation state.
Binding of integrin receptors to extracellular ligands is a complex process involving receptor-ligand interactions at the cell-substrate interface, signals activating the receptors, and assembly of cytoskeletal and adhesion plaque proteins at the cytoplasmic face. To analyze the contribution of these elements to overall cell adhesion, we have developed a model system that characterizes the functional binding characteristic for adhesion receptors as the force required to separate the integrin-ligand bond. A spinning disk device was used to apply a range of controlled hydrodynamic forces to adherent cells. The adhesion of K562 erythroleukemia cells, a cell line expressing a single fibronectin receptor, integrin alpha5beta1, which was uniformly activated with the monoclonal antibody TS2/16, to defined fibronectin surface densities was examined. Cell adhesion strength increased linearly with receptor and ligand densities. Based on chemical equilibrium principles, it is shown that adhesion strength is directly proportional to the number of receptor-ligand bonds. This analysis provides for the definition of a new physical parameter, the adhesion constant psi, which is related to the bond strength and binding equilibrium constant and has units of force-length2. This parameter can be measured by the experimental system presented and is governed by the activation state of integrin receptors. This simplified model isolates the integrin receptor-ligand binding parameters and provides a basis for analysis of the functions of signaling and cytoskeletal elements in the adhesion process.
García
A J
AJ
Department of Microbiology, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA. garciaa@mail.med.upenn.edu
Huber
F
F
Boettiger
D
D
D002448:Cell Adhesion; D005353:Fibronectins / Q000378:metabolism*; D006801:Humans; D016023:Integrins / Q000378:metabolism*; D008024:Ligands; D008954:Models, Biological; D011956:Receptors, Cell Surface / Q000378:metabolism; D019030:Receptors, Vitronectin; D014407:Tumor Cells, Cultured
D005353:Fibronectins; D016023:Integrins; D008024:Ligands; D011956:Receptors, Cell Surface; D019030:Receptors, Vitronectin; C094340:integrin alphavbeta1