C-terminal opening mimics 'inside-out' activation of integrin alpha5beta1.
Integrins are adhesion molecules that convey signals both to and from the cytoplasm across the plasma membrane. In resting cells, integrins in a low affinity state can be activated by 'inside-out signaling', in which signals affecting integrin heterodimer cytoplasmic domains cause a conformational change in the integrin ligand-binding headpiece connected to the membrane by two long, approximately 16 nm stalks. Here we demonstrate a mechanism for conveying a conformational change over the long distance from the plasma membrane to the headpiece. We prepared soluble, alpha5beta1 integrin heterodimer extracellular fragments in which interactions between alpha- and beta-subunit cytoplasmic domains were replaced with an artificial clasp. Release of this C-terminal clasp by specific protease cleavage resulted in an approximately 14 nm separation of the stalks coupled to increased binding to fibronectin. This activation did not require any associated molecules or clustering and was observed with physiological concentrations of divalent cations. These findings suggest that the overall mechanism for integrin inside-out activation involves the spatial separation of the cytoplasmic and/or transmembrane domains.
Takagi
J
J
The Center for Blood Research, Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115, USA. takagi@cbr.med.harvard.edu
Erickson
H P
HP
Springer
T A
TA
D000595:Amino Acid Sequence; D002413:Cations, Divalent / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D003593:Cytoplasm / Q000737:chemistry / Q000378:metabolism; D019281:Dimerization; D010450:Endopeptidases / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D008024:Ligands; D008854:Microscopy, Electron; D008954:Models, Biological; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D021122:Protein Subunits; D017469:Receptors, Fibronectin / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D015398:Signal Transduction; D012995:Solubility
D002413:Cations, Divalent; D005353:Fibronectins; D008024:Ligands; D010446:Peptide Fragments; D021122:Protein Subunits; D017469:Receptors, Fibronectin; D010450:Endopeptidases; C099488:TEV protease