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- W2010704526 abstract "The platelet integrin αIIbβ3 plays a key role in platelet adhesion, activation, and aggregation at the subendothelium and at protein-coated synthetic biomaterials. In this study, interactions between αIIbβ3 and both protein and peptide ligands for the receptor were imaged under physiological conditions by high-resolution atomic force microscopy (AFM). To directly image the ligand−receptor interactions, αIIbβ3 receptors were reconstituted into a supported lipid bilayer formed on a mica surface in the AFM fluid cell assembly and subsequently activated with Mn2+. Fibrinogen, the natural protein ligand for the integrin, as well as a nanogold-labeled peptide ligand (an RGD-containing heptamer) were infused into the AFM fluid cell, incubated with the reconstituted and activated receptors, and imaged under buffer. Height images illustrating topographical features showed the integrin reconstituted in the bilayer. Fibrinogen molecules binding to the receptors were easily observed in the height images, with fibrinogen showing its characteristic trinodular structure and occasionally bridging integrin receptors. Fibrinogen was observed to bind to integrins at the D-domain consistent with the location of the γ-chain dodecapeptide, while fibrinogen bridging integrins bound to receptors on opposite sides of the protein consistent with a 2-fold axis of symmetry. Peptide ligands were not visible in height images; however, phase images that map the mechanical properties detected the nanogold labels and demonstrated the presence of peptide ligands bound to the receptors. The results demonstrate the ability of this high-resolution microscopy technique to directly visualize single ligand/receptor interactions in a dynamic and physiologically relevant environment, and establish a framework for future fundamental studies of single protein/receptor interactions during normal pathological processes as well as biomaterial surface-induced thrombosis." @default.
- W2010704526 created "2016-06-24" @default.
- W2010704526 creator A5031765084 @default.
- W2010704526 creator A5032923334 @default.
- W2010704526 creator A5063690163 @default.
- W2010704526 date "2005-06-23" @default.
- W2010704526 modified "2023-09-25" @default.
- W2010704526 title "AFM Imaging of Ligand Binding to Platelet Integrin α<sub>IIb</sub>β<sub>3</sub> Receptors Reconstituted into Planar Lipid Bilayers" @default.
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- W2010704526 doi "https://doi.org/10.1021/la046943h" @default.
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