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

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• 作者：Kabir H. Biswas ; Jay T. Groves
• 刊名：Langmuir
• 出版年：2016
• 出版时间：July 5, 2016
• 年：2016
• 卷：32
• 期：26
• 页码：6775-6780
• 全文大小：367K
• 年卷期：0
• ISSN：1520-5827

文摘

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.