Visualization of perforin/gasdermin/complement-formed pores in real cell membranes using atomic force microscopy

Cell Mol Immunol. 2019 Jun;16(6):611-620. doi: 10.1038/s41423-018-0165-1. Epub 2018 Oct 3.

Abstract

Different types of pores ubiquitously form in cell membranes, leading to various types of cell death that profoundly influence the fate of inflammation and the disease status. However, these pores have never truly been visualized to date. Atomic force microscopy (AFM), which is emerging as a powerful tool to analyze the mechanical properties of biomolecules and cells, is actually an excellent imaging platform that allows biological samples to be visualized by probing surface roughness at the level of atomic resolution. Here, membrane pore structures were clearly visualized using AFM. This visualization not only describes the aperture and depth of the pore complexes but also highlights differences among the pores formed by perforin and gasdermins in tumor cell membranes and by complement in immune cell membranes. Additionally, this type of visualization also reveals the dynamic process of pore formation, fusion, and repair.

Keywords: AFM; GSDMD/GSDME; complement; membrane pore formation; perforin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins
  • CD8-Positive T-Lymphocytes / immunology*
  • Cell Membrane / metabolism*
  • Cell Membrane / ultrastructure
  • Cells, Cultured
  • Complement System Proteins / metabolism*
  • Cytotoxicity, Immunologic
  • Immunological Synapses / ultrastructure
  • Melanoma, Experimental
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Atomic Force / methods*
  • Neoplasm Proteins / metabolism*
  • Perforin / metabolism*
  • Pore Forming Cytotoxic Proteins / metabolism*
  • Streptolysins

Substances

  • Bacterial Proteins
  • Gsdma protein, mouse
  • Neoplasm Proteins
  • Pore Forming Cytotoxic Proteins
  • Streptolysins
  • streptolysin O
  • Perforin
  • Complement System Proteins