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      A Microbead Supported Membrane-Based Fluorescence Imaging Assay Reveals Intermembrane Receptor-Ligand Complex Dimension with Nanometer Precision.

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          Abstract

          Receptor-ligand complexes spanning a cell-cell interface inevitably establish a preferred intermembrane spacing based on the molecular dimensions and orientation of the complexes. This couples molecular binding events to membrane mechanics and large-scale spatial organization of receptors on the cell surface. Here, we describe a straightforward, epi-fluorescence-based method to precisely determine intermembrane receptor-ligand dimension at adhesions established by receptor-ligand binding between apposed membranes in vitro. Adhesions were reconstituted between planar and silica microbead supported membranes via specific interaction between cognate receptor/ligand pairs (EphA2/EphrinA1 and E-cadherin/anti-E-cadherin antibody). Epi-fluorescence imaging of the ligand enrichment zone in the supported membrane beneath the adhering microbead, combined with a simple geometrical interpretation, proves sufficient to estimate intermembrane receptor-ligand dimension with better than 1 nm precision. An advantage of this assay is that no specialized equipment or imaging methods are required.

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          Author and article information

          Journal
          Langmuir
          Langmuir : the ACS journal of surfaces and colloids
          American Chemical Society (ACS)
          1520-5827
          0743-7463
          July 05 2016
          : 32
          : 26
          Affiliations
          [1 ] Mechanobiology Institute, National University of Singapore , Singapore 117411, Singapore.
          [2 ] Department of Chemistry, University of California , Berkeley, California 94720, United States.
          Article
          10.1021/acs.langmuir.6b01377
          27264296

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