title: Two-stage activation for alpha5beta1 integrin binding to surface-adsorbed fibronectin.
abstract: By analyzing the functional binding of alpha5beta1 integrin to adsorbed fibronectin in intact cells, we demonstrate that integrin activation results in linear increases in adhesion strength as a function of ligand density, suggesting that modulation of the receptor-ligand interaction is the dominant mechanism for adhesion during the initial stages of adhesion and that cooperative binding contributes little to initial adhesion strength. Using this experimental framework, we show the existence of three distinct activation states for alpha5beta1 integrin binding to adsorbed fibronectin for both passive, antibody-induced and active, cell-controlled activation. During the initial phase of adhesion, alpha5beta1 integrin is activated in an energy-dependent process from the nonbinding ground state to an intermediate state in which the receptor binds fibronectin and provides significant mechanical coupling. In later stages of adhesion maturation, alpha5beta1 integrin is activated to a higher binding state, which provides significant increases in adhesion strength compared with the intermediate state. These multiple binding states most likely result from different integrin conformations and reflect distinct interactions between alpha5beta1 and sites on adsorbed fibronectin. Multiple activation states for alpha5beta1 suggest the existence of distinct stages in adhesion signaling and strengthening and can provide a versatile mechanism for the regulation of adhesive interactions.
authors.affiliation: Department of Microbiology, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
authors.forename: A J
authors.initials: AJ
authors.lastname: García
authors.forename: J
authors.initials: J
authors.lastname: Takagi
authors.forename: D
authors.initials: D
authors.lastname: Boettiger
mesh_terms: D000327:Adsorption; D002448:Cell Adhesion; D016023:Integrins / Q000378:metabolism*; D011485:Protein Binding; D017469:Receptors, Fibronectin / Q000378:metabolism*
chemical_list: D016023:Integrins; D017469:Receptors, Fibronectin