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Related Experiment Videos

Two proteins targeted to the same lytic granule compartment undergo very different posttranslational processing.

J K Burkhardt1, S Hester, Y Argon

  • 1Department of Microbiology and Immunology, Duke University Medical Center, Durham, NC 27710.

Proceedings of the National Academy of Sciences of the United States of America
|September 1, 1989
PubMed
Summary

Natural killer (NK) cells release cytolysin and serine proteases from granules. These proteins undergo distinct post-translational modifications, suggesting unique targeting mechanisms for NK cell granule packaging.

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Area of Science:

  • Immunology
  • Cell Biology

Background:

  • Natural killer (NK) cells are crucial for innate immunity, utilizing cytotoxic granules to eliminate target cells.
  • These granules contain effector proteins like cytolysin and serine proteases, essential for cell-mediated cytotoxicity.

Purpose of the Study:

  • To investigate the subcellular localization of cytolysin and serine proteases within NK cell granules.
  • To analyze the post-translational modifications of these proteins and infer their intracellular targeting pathways.

Main Methods:

  • Immunofluorescence microscopy was employed to visualize protein distribution.
  • Immunoelectron microscopy provided high-resolution localization within granule compartments.
  • Analysis of N-linked oligosaccharide processing and phosphomannose modifications was performed.

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Main Results:

  • Both cytolysin and serine proteases (including granzyme A) were localized to the core regions of dense core granules.
  • Cytolysin exhibited complex-type N-linked oligosaccharides, while granzyme A had high-mannose-type oligosaccharides.
  • Granzyme A, but not cytolysin, was modified with phosphomannose moieties.

Conclusions:

  • NK cell granule proteins are specifically targeted to the granule core.
  • The distinct post-translational modifications suggest differential protein sorting mechanisms.
  • The mannose 6-phosphate pathway may be involved, but cytolysin's processing indicates an alternative NK granule targeting system.