brij-58 and Glioma

This paper describes a simple and direct procedure for assaying Ca(2+)-dependent protein kinase C (PKC) activity in membrane fractions isolated from purified murine B lymphocytes (B cells) treated with phorbol 12-myristate 13-acetate (PMA). The results indicate that membrane-bound PKC in B cells, treated with PMA, can be measured directly in the presence of 0.5% Brij 58 by assaying the transfer of 32P from [gamma-32P]ATP to histone type III-S. This method obviates the need for partial purification of the protein kinase by ion-exchange chromatography prior to assaying PKC activity. The properties of membrane-associated PKC activity in B cells have been characterized, and the kinetics of PMA-induced translocation of PKC in cultured murine B cells, the rat glial tumor clone C6, and primary neonatal osteoblastic cells have been defined by this direct assay. The results obtained with B cells and the other cell lines indicate that this direct assay procedure could be useful for studies on the factors controlling PKC translocation in a variety of cultured mammalian cells.

Topics: Adenosine Triphosphate; Animals; B-Lymphocytes; Cell Fractionation; Cell Membrane; Cells, Cultured; Cetomacrogol; Cytosol; Detergents; Female; Glioma; Kinetics; Mice; Mice, Inbred DBA; Osteoblasts; Phosphorus Radioisotopes; Protein Kinase C; Radioisotope Dilution Technique; Rats; Tetradecanoylphorbol Acetate; Tumor Cells, Cultured