Divalent cations regulate the folding and activation status of integrins during their intracellular trafficking

Integrins are divalent cation-dependent, alpha beta heterodimeric adhesion receptors that control many fundamental aspects of cell behaviour by bi-directional signalling between the extracellular matrix and intracellular cytoskeleton. The activation state of cell surface integrins is tightly regulated by divalent cation occupancy of the ligand-binding pocket and by interaction with cytoplasmic adaptor proteins, such as talin. These agents elicit gross conformational changes across the entire molecule, which specify the activation state. Much less is known about the activation state of newly synthesised integrins or the role of cations during the early folding and trafficking of integrins. Here we use a number of well-characterised, conformation-specific antibodies to demonstrate that beta 1-integrins adopt the bent, inactive conformation after assembly with beta-integrins in the endoplasmic reticulum. Folding and assembly are totally dependent on the binding of Ca2+ ions. In addition, Ca2+ binding prevents integrin activation before its arrival at the cell surface. Activation at the cell surface occurs only following displacement of Ca2+ with Mg2+ or Mn2+. These results demonstrate the essential roles played by divalent cations to facilitate folding of the beta-integrin subunit, to prevent inappropriate intracellular integrin signalling, and to activate ligand binding and signalling at the cell surface.