Activation of protein kinase C in neutrophil cytoplasts

FEBS Letters

Gennaro, Renato¹  Romeo, Domenico¹  Florio, Chiara¹ 

[1] Istituto di Chimica Biologica dell'Università, 34127 Trieste, Italy

关键词: Neutrophil cytoplast;    Protein phosphorylation;    Protein kinase C;    Superoxide anion;    Phorbol myristate acetate;    PMA;    phorbol 12-O-myristate-13acetate;    TFP;    trifluoperazine;    OAG;    1-oleyl-2-acetylglycerol;    PBS;    phosphate-buffered saline;    SDS-PAGE;    sodium dodecyl sulfate-polyacrylamide gel electrophoresis;    B12BP;    vitamin B12 binding protein;   

DOI  :  10.1016/0014-5793(85)81068-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Treatment of enucleated, granule-free neutrophil cytoplasts with the protein kinase C activator phorbol 12O-myristate-13-acetate (PMA) causes an increased -³²P-incorporation into a variety of polypeptides. Permeabilization of PMA-stimulated, ³²P-labeled cytoplasts by 0.01% digitonin fully releases the majority of these phosphorylated proteins. A statistically significant correlation is found between the extent of PMA-induced activation of generation of Superoxide anion (O⁻ ₂) and the phosphorylation of a cytosolic polypeptide with an apparent M _r, of 46000, whose -³²P-labeling is also enhanced by the treatment of cytoplasts with 1-oleyl-2-acetylglycerol, the Ca²⁺ ionophore ionomycin or latex beads. Furthermore, treatment of cytoplasts with the protein kinase C inhibitor trifluoperazine markedly inhibits the ³²P-labeling of proteins in the 40000 M _r range, including the 46 kDa polypeptide, and almost totally abolishes the activation of O⁻ ₂ production by PMA.

【 授权许可】

Unknown