P-selectin mobility undergoes a sol-gel transition as it diffuses from exocytosis sites into the cell membrane
journal contributionposted on 2022-06-06, 13:41 authored by Nicola Hellen, Gregory I Mashanov, Ianina L Conte, Sophie le Trionnaire, Victor Babich, Laura Knipe, Alamin Mohammed, Kazim Ogmen, Silvia Martin-Almedina, Katalin Török, Matthew J Hannah, Justin E Molloy, Tom Carter
In response to vascular damage, P-selectin molecules are secreted onto the surface of cells that line our blood vessels. They then serve as mechanical anchors to capture leucocytes from the blood stream. Here, we track individual P-selectin molecules released at the surface of live endothelial cells following stimulated secretion. We find P-selectin initially shows fast, unrestricted diffusion but within a few minutes, movement becomes increasingly restricted and ~50% of the molecules become completely immobile; a process similar to a sol-gel transition. We find removal of the extracellular C-type lectin domain (ΔCTLD) and/or intracellular cytoplasmic tail domain (ΔCT) has additive effects on diffusive motion while disruption of the adapter complex, AP2, or removal of cell-surface heparan sulphate restores mobility of full-length P-selectin close to that of ΔCT and ΔCTLD respectively. We have found P-selectin spreads rapidly from sites of exocytosis and evenly decorates the cell surface, but then becomes less mobile and better-suited to its mechanical anchoring function.