Translocation of proteins homologous to human neutrophil p47phox and p67phox to the cell membrane in activated hemocytes of Galleria mellonella.
Activation of the superoxide forming respiratory burst oxidase of human neutrophils, crucial in host defence, requires the cytosolic proteins p47phox and p67phox which translocate to the plasma membrane upon cell stimulation and activate flavocytochrome b558, the redox centre of this enzyme system. We have previously demonstrated the presence of proteins (67 and 47kDa) in hemocytes of the insect Galleria mellonella homologous to proteins of the superoxide-forming NADPH oxidase complex of neutrophils. The work presented here illustrates for the first time translocation of homologous hemocyte proteins, 67 and 47kDa from the cytosol to the plasma membrane upon phorbol 12-myristate 13 acetate (PMA) activation. In hemocytes, gliotoxin (GT), the fungal secondary metabolite significantly suppressed PMA-induced superoxide generation in a concentration dependent manner and reduced translocation to basel nonstimulated levels. Primarily these results correlate translocation of hemocyte 47 and 67kDa proteins with PMA induced oxidase activity. Collectively results presented here, demonstrate further cellular and functional similarities between phagocytes of insects and mammals and further justify the use of insects in place of mammals for modelling the innate immune response to microbial pathogens.
Renwick
Julie
J
Medical Mycology Unit, National Institute for Cellular Biotechnology, Department of Biology, NUI Maynooth, Co. Kildare, Ireland.
Reeves
Emer P
EP
Wientjes
Frans B
FB
Kavanagh
Kevin
K
D000818:Animals; D002462:Cell Membrane / Q000201:enzymology* / Q000276:immunology; D002478:Cells, Cultured; D005912:Gliotoxin / Q000494:pharmacology; D006434:Hemocytes / Q000187:drug effects / Q000201:enzymology* / Q000276:immunology; D006801:Humans; D009036:Moths / Q000187:drug effects / Q000201:enzymology* / Q000276:immunology; D019255:NADPH Oxidases / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D021381:Protein Transport / Q000276:immunology; D016897:Respiratory Burst / Q000276:immunology; D017386:Sequence Homology, Amino Acid
D010750:Phosphoproteins; C079789:neutrophil cytosol factor 67K; D005912:Gliotoxin; D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1