CX3CL1/Fractalkine acts as a ligand for both CX3CR1 and integrins. It binds to CX3CR1 and to integrins ITGAV:ITGB3 and ITGA4:ITGB1. It can activate integrins in both a CX3CR1-dependent and CX3CR1-independent manner. In the presence of CX3CR1, it activates integrins by binding to the classical ligand-binding site (site 1) in integrins. In the absence of CX3CR1, it binds to a second site (site 2) in integrins which is distinct from site 1 and enhances the binding of other integrin ligands to site 1.

It soluble form is chemotactic for T-cells and monocytes and not for neutrophils. The membrane-bound form promotes adhesion of those leukocytes to endothelial cells. Investigators believe it may play a role in regulating leukocyte adhesion and migration processes at the endothelium.