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Related Experiment Videos

Nucleosomes bind to cell surface proteoglycans.

K Watson1, N J Gooderham, D S Davies

  • 1Section on Clinical Pharmacology, Imperial College School of Medicine, Hammersmith Hospital, DuCane Road, London W12 0NN, United Kingdom. kwatson@rpms.ac.uk

The Journal of Biological Chemistry
|July 27, 1999
PubMed
Summary
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Sulfated polysaccharides like dextrin 2-sulfate bind to nucleosomes on activated T-cells. This interaction, mediated by histone N termini and heparan sulfate proteoglycans, reveals a novel cell surface complex.

Area of Science:

  • Immunology
  • Cell Biology
  • Biochemistry

Background:

  • Activated T-cells exhibit surface-associated nucleosomes.
  • Nucleosomes are DNA-protein complexes typically found within the nucleus.

Purpose of the Study:

  • To investigate the interaction between sulfated polysaccharides and cell surface nucleosomes on activated T-cells.
  • To elucidate the mechanism of nucleosome anchoring to the T-cell plasma membrane.

Main Methods:

  • Incubation of activated T-cells with dextrin 2-sulfate (D2S).
  • Purification of cell surface material using anion-exchange chromatography.
  • Characterization of purified complexes via DNA fragmentation analysis and antibody binding assays.
  • Investigation of sulfated polysaccharide-nucleosome interaction using biotinylated D2S.

Related Experiment Videos

  • Enzymatic treatment of T-cells with heparinase/heparitinase.

    • Main Results:

    • Dextrin 2-sulfate displaced nucleosomes (histones H2A, H2B, H3, H4 complexed with DNA) from activated T-cell surfaces.
    • Purified nucleosomes showed DNA fragmentation patterns similar to apoptotic cells.
    • An antibody against nucleosomes confirmed their presence on the T-cell plasma membrane.
    • Sulfated polysaccharides bind to nucleosomes via histone N-termini (H2A, H2B).
    • Heparinase/heparitinase treatment abolished nucleosome binding, indicating heparan sulfate proteoglycan involvement.
    • Nucleosomes are anchored by heparan sulfate proteoglycans via ionic interactions with histone N-termini.
    • Bound nucleosomes can further bind other sulfated polysaccharides, forming a ternary complex.

    Conclusions:

    • Nucleosomes are present on the surface of activated T-cells.
    • Heparan sulfate proteoglycans anchor nucleosomes to the T-cell membrane through ionic interactions with histone N-termini.
    • This interaction facilitates the binding of additional sulfated polysaccharides, forming a complex on the cell surface.