Protein phosphorylation in alveolar macrophages after stimulation with heavy metal-coated silica particles.
The aim of our study was to determine agonist- and time-dependent phosphorylation of distinct proteins in bovine alveolar macrophages after activation with quartz dusts and heavy metal-coated silica particles. Silica particles were coated with vanadium oxide or cadmium oxide. Separation of proteins was achieved by one-dimensional and optimized horizontal two-dimensional polyacrylamide gel electrophoresis. Protein phosphorylation was either determined by [32P] labeling and subsequent autoradiography or Western blotting using monoclonal antibodies against phosphotyrosine. One of the phosphorylated proteins was identified as the heat shock protein HSP-27. Stimulation with cadmium oxide-coated silica particles and PMA led to a significant increase of phosphorylated HSP-27. Additionally cadmium oxide-coated silica particles induced tyrosine phosphorylation of six main proteins. Stimulation with vanadium oxide-coated silica particles drastically increased the amount of tyrosine phosphorylated proteins while this effect was not observed with uncoated silica particles.
Radloff
M
M
Department of Biochemistry and Molecular Biology, University of Hamburg, Germany.
Delling
M
M
Gercken
G
G
D000818:Animals; D019187:Cadmium Compounds / Q000633:toxicity*; D002417:Cattle; D004391:Dust; D006360:Heat-Shock Proteins / Q000378:metabolism*; D007700:Kinetics; D008262:Macrophage Activation / Q000187:drug effects; D016676:Macrophages, Alveolar / Q000187:drug effects* / Q000378:metabolism*; D010087:Oxides / Q000633:toxicity*; D010766:Phosphorylation / Q000187:drug effects; D011791:Quartz / Q000633:toxicity; D020794:Receptor Protein-Tyrosine Kinases / Q000187:drug effects; D012822:Silicon Dioxide / Q000633:toxicity*; D013268:Stimulation, Chemical; D014443:Tyrosine / Q000378:metabolism; D017968:Vanadium Compounds / Q000633:toxicity*
D019187:Cadmium Compounds; D004391:Dust; D006360:Heat-Shock Proteins; D010087:Oxides; D017968:Vanadium Compounds; C029663:cadmium oxide; D011791:Quartz; D014443:Tyrosine; D012822:Silicon Dioxide; C066075:vanadium pentoxide; D020794:Receptor Protein-Tyrosine Kinases