Quantification of membrane protein diffusion on endothelial cell surfaces under shear stress using a photochromic fluorescent protein

DOI Web Site 参考文献24件
  • Kawasaki Naoto
    Graduate School of Engineering of Functional Control Systems, Shibaura Institute of Technology
  • Hamasaki Keita
    Department of Applied Chemistry, Shibaura Institute of Technology
  • Sasaki Saori
    Department of Mechanical Engineering, Kyushu University
  • Takeishi Naoki
    Department of Mechanical Engineering, Kyushu University
  • Kudo Susumu
    Department of Mechanical Engineering, Kyushu University

説明

<p>The fluidity of endothelial cell (EC) membranes, consisting of a lipid bilayer and heterogeneous multicomponent, undergoes alterations in response to shear stress. Although localized variations in membrane fluidity are assumed to induce region-specific signal transduction by modulating membrane protein dynamics on almost planar bilayers, membrane protein diffusion under shear stress remains uncertain. Hence, this study aimed to quantify membrane protein diffusion on ECs under fluid shear stress. We used the photochromic fluorescent protein Dronpa to tag a glycosylphosphatidylinositol-anchored protein (GPI-AP), which diffuses across the outer membranes, and quantified its surface diffusion based on the spatiotemporal distribution of Dronpa-Green-labeled GPI-AP (DGGPI-AP) on EC membranes. We developed an experimental platform to measure the GPI-AP surface diffusion under fluid shear stress and quantified the diffusion coefficient of GPI-AP in two distinct membrane regions: upstream and downstream relative to the direction of fluid flow. Our experimental results showed that there were not statistically significant differences in GPI-AP diffusion on EC membranes between the time points or between the upstream and downstream regions of ECs for at least 15 minutes under shear stress. Our developed methodology and experimental results will be useful to understand a relationship between the membrane protein diffusion and shear-induced cellular processes.</p>

収録刊行物

  • Journal of Biorheology

    Journal of Biorheology 38 (2), 88-93, 2024

    特定非営利活動法人 日本バイオレオロジー学会

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