Subject(s)
Cell Communication , Erythrocytes/physiology , Fibroblasts/physiology , Membrane Proteins/biosynthesis , Proteins/metabolism , Animals , Cell Membrane/physiology , Coculture Techniques , Culture Media, Conditioned/toxicity , Erythrocytes/cytology , Fibroblasts/cytology , Humans , K562 Cells , Membrane Proteins/toxicity , Mice , Proteins/toxicitySubject(s)
Cytokines/biosynthesis , Killer Cells, Lymphokine-Activated/metabolism , Apoptosis , Blotting, Western , Calcimycin/pharmacology , Cell Line , Cytokines/immunology , Electrophoresis, Polyacrylamide Gel , Humans , Immunoenzyme Techniques , Interleukin-2/pharmacology , Killer Cells, Lymphokine-Activated/drug effectsSubject(s)
Cyclic AMP-Dependent Protein Kinases/physiology , Cytotoxicity, Immunologic , Killer Cells, Lymphokine-Activated/physiology , Protein Kinase C/physiology , Serine Endopeptidases/physiology , Animals , Apoptosis , Bucladesine/pharmacology , Granzymes , Humans , L Cells , Mice , Tetradecanoylphorbol Acetate/pharmacology , Tumor Cells, Cultured , Tumor Necrosis Factor-alpha/pharmacologySubject(s)
Apoptosis/immunology , Blood Proteins/immunology , Cytotoxicity, Immunologic , Killer Cells, Lymphokine-Activated/immunology , Ammonium Chloride/pharmacology , Calcium/antagonists & inhibitors , DNA/metabolism , Egtazic Acid/pharmacology , Humans , Hydrolysis , Lysosomes/drug effects , Lysosomes/enzymology , Necrosis , Tumor Cells, CulturedABSTRACT
The catalytic subunit of cAMP-dependent protein kinase modifies proteins of NKCF. Several proteins of the treated NKCF can be labelled with 32P. Treatment of NKCF with protein kinase enhance the cytotoxicity of NKCF.
Subject(s)
Cytotoxicity, Immunologic , Killer Cells, Natural/immunology , Protein Kinases/metabolism , Proteins/immunology , Catalysis , Humans , Killer Cells, Natural/metabolism , Killer Factors, Yeast , Phosphorylation , Proteins/metabolismABSTRACT
The protein composition of the cytolytic factor (NKCF) secreted by natural killer cells was analyzed. Natural killer cells were isolated from spleens of nude rats and purified in the Percole gradient. SDS-PAAG analysis of NKCF revealed the presence of major proteins with an approximate molecular weight of 62-67 kDa. Biochemical properties of these proteins such as fractionation on a TSK-3000 SW gel filtration column, electrophoretical mobility in reduced conditions, enhanced cytolytical activity in the presence of Ca2+ demonstrate the similarity with the pore-forming protein--perforin. Apart from the major component, proteins with m. w. 42 kDa, 31-28 kDa, 17 kDa and 12-14 kDa were also found. It is possible that the Ca2+-independent cytolytical activity is bound to one of these minor protein components of NKCF.