Recruitment of actin to the cytoskeletons of human monocyte-like cells activated by complement fragment C5a. Is protein kinase C involved?
U-937 cells differentiated by exposure to dibutyryl cyclic AMP respond to complement fragment C5a with a marked increase in cytoskeletal F-actin, which can be detected by fluorescence-activated cell sorting (f.a.c.s.) analysis of their rhodamine phalloidin-stained cytoskeletons. The C5a-induced increase in F-actin content can be prevented by prior exposure of the cells to cytochalasin B and pertussis toxin. It is insensitive to removal of extra cellular Ca2+, to cholera toxin or to neomycin. Phorbol myristate acetate (PMA), an activator of protein kinase C, does not induce actin polymerization in the differentiated cells. Both C5a and PMA stimulate superoxide production. The action of C5a on superoxide formation is also inhibited by neomycin, a phospholipase inhibitor. These results suggest that the cytoskeletal response to C5a requires activation of a G protein, but probably does not involve phospholipase C and protein kinase C, and is not highly dependent on the availability of Ca2+. Phospholipase C and kinase C may, however, be components of the pathway leading from C5a binding to superoxide production.
Department of Biochemistry, University of Sheffield, U.K.
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Banks
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Barker
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Burton
D000199:Actins / Q000378:metabolism*; D001427:Bacterial Toxins / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D002460:Cell Line; D003182:Complement C5 / Q000378:metabolism*; D015936:Complement C5a; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D008856:Microscopy, Fluorescence; D009000:Monocytes / Q000187:drug effects / Q000378:metabolism*; D009355:Neomycin / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000199:Actins; D001427:Bacterial Toxins; D003182:Complement C5; D015936:Complement C5a; D011493:Protein Kinase C; D009355:Neomycin; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium