Cho J, Mosher D F
Department of Medicine, Molecular and Cellular Pharmacology Program, University of Wisconsin-Madison School of Medicine, 1300 University Avenue, Madison, WI 53706, USA.
J Thromb Haemost. 2006 May;4(5):943-51. doi: 10.1111/j.1538-7836.2006.01862.x.
Various types of laminin (LN) are ubiquitous components of basement membrane and exposed to blood upon localized damage of vascular endothelial cells. Fibronectin is a plasma protein that is insolubilized into fibrils in a regulated fashion by, for example, lysophosphatidic acid (LPA)-stimulated fibroblasts or platelets spread on supportive adhesive ligands.
To study assembly of plasma fibronectin by LPA-activated platelets adherent to LN-111 via alpha6beta1 integrin.
Platelets adherent to LN-111-bound plasma fibronectin or its N-terminal 70 kD fragment in fibrillar arrays at the periphery of spread platelets under static but not shear conditions. Formation of fibronectin arrays under static conditions was inhibited by co-incubation with the N-terminal 70 kD fragment or with a 49-amino acid peptide that binds to the N-terminal region of fibronectin. Approximately 7000 fibronectin dimers bound per adherent platelet with a K(d) of 50 nm. Bound 70 kD fragment was readily solubilized with deoxycholate (DOC), whereas bound fibronectin became progressively insoluble. Bound 70 kD fragment became resistant to DOC extraction after treatment with a cell-impermeable, reducible crosslinker. Crosslinked 70 kD fragment was found in a high molecular weight complex. As with fibroblasts, signaling molecules modulating actin cytoskeletal organization controlled expression of binding sites for the N-terminal 70 kD region of fibronectin on adherent platelets.
These results indicate that platelets adherent to LN-111 via alpha(6)beta(1) support subsequent assembly of fibronectin, but possibly only under conditions of intermittent or stagnant blood flow.
