Label-free analysis of membrane protein binding kinetics and cell adhesions using evanescent scattering microscopy.

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  • Additional Information
    • Source:
      Publisher: Royal Society of Chemistry Country of Publication: England NLM ID: 0372652 Publication Model: Electronic Cited Medium: Internet ISSN: 1364-5528 (Electronic) Linking ISSN: 00032654 NLM ISO Abbreviation: Analyst Subsets: MEDLINE
    • Publication Information:
      Publication: Cambridge : Royal Society of Chemistry
      Original Publication: London : Chemical Society
    • Subject Terms:
      Membrane Proteins* ; Microscopy*/methods; Cell Adhesion ; Ligands ; Protein Binding ; Kinetics
    • Abstract:
      Measuring ligand interactions with membrane proteins in single live cells is critical for understanding many cellular processes and screening drugs. However, developing such a capability has been a difficult challenge. Here, we employ evanescent scattering microscopy (ESM) to show that ligand binding to membrane proteins can change the cell adhesion properties, which are intrinsic cell properties and independent of random cell micromotions and ligand mass, thus allowing the kinetics analyses of both proteins and small molecules binding to membrane proteins in both single fixed and live cells. In addition, utilizing the high spatiotemporal resolution of ESM, the positions of cell adhesion sites can be tracked in real-time to analyze the cell deformations and migrations, thus providing a potential approach for understanding the cell activity during the ligand binding process in detail. The presented method may pave the road for developing a versatile and easy-to-use label-free detection strategy for in situ analysis of molecular interaction dynamics in living biosystems with single-cell resolution.
    • Accession Number:
      0 (Membrane Proteins)
      0 (Ligands)
    • Publication Date:
      Date Created: 20230906 Date Completed: 20231006 Latest Revision: 20231006
    • Publication Date:
      20231215
    • Accession Number:
      10.1039/d3an00977g
    • Accession Number:
      37671903