Membrane receptors are not required to deliver granzyme B during killer cell attack.
Granzyme B (GzmB), a serine protease of cytotoxic T lymphocytes and natural killer (NK) cells, induces apoptosis by caspase activation after crossing the plasma membrane of target cells. The mechanism of this translocation during killer cell attack, however, is not understood. Killer cells release GzmB and the membrane-disturbing perforin at the contact site after target recognition. Receptor-mediated import of glycosylated GzmB and release from endosomes were suggested, but the role of the cation-independent mannose 6-phosphate receptor was recently refuted. Using recombinant nonglycosylated GzmB, we observed binding of GzmB to cellular membranes in a cell type-dependent manner. The basis and functional impact of surface binding were clarified. GzmB binding was correlated with the surface density of heparan sulfate chains, was eliminated on treatment of target cells with heparinase III or sodium chlorate, and was completely blocked by an excess of catalytically inactive GzmB or GzmK. Although heparan sulfate-bound GzmB was taken up rapidly into intracellular lysosomal compartments, neither of the treatments had an inhibitory influence on apoptosis induced by externally added streptolysin O and GzmB or by natural killer cells. We conclude that membrane receptors for GzmB on target cells are not crucial for killer cell-mediated apoptosis.
Kurschus
Florian C
FC
Department of Neuroimmunology, Max-Planck-Institute of Neurobiology, Am Klopferspitz 18, D-82152 Planegg-Martinsried, Germany. kurschus@neuro.mpg.de
Bruno
Roxana
R
Fellows
Edward
E
Falk
Christine S
CS
Jenne
Dieter E
DE
D017209:Apoptosis; D001665:Binding Sites; D002460:Cell Line; D002463:Cell Membrane Permeability; D053804:Granzymes; D006493:Heparin / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism; D006801:Humans; D007694:Killer Cells, Natural / Q000378:metabolism / Q000502:physiology*; D008247:Lysosomes / Q000378:metabolism; D011509:Proteoglycans / Q000032:analysis / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000502:physiology*; D012697:Serine Endopeptidases / Q000378:metabolism*
D011509:Proteoglycans; D011956:Receptors, Cell Surface; C036090:heparin proteoglycan; D006493:Heparin; C509213:GZMB protein, human; D053804:Granzymes; D012697:Serine Endopeptidases