A putative link between phagocytosis-induced apoptosis and hemocyanin-derived phenoloxidase activation.
Apoptosis and phagocytosis are crucial processes required for developmental morphogenesis, pathogen deterrence and immunomodulation in metazoans. We present data showing that amebocytes of the chelicerate, Limulus polyphemus, undergo phagocytosis-induced cell death after ingesting spores of the fungus, Beauveria bassiana, in vitro. The observed biochemical and morphological modifications associated with dying amebocytes are congruent with the hallmarks of apoptosis, including: extracellularisation of phosphatidylserine, intranucleosomal DNA fragmentation and an increase in caspase 3/7-like activities. Previous studies have demonstrated that phosphatidylserine is a putative endogenous activator of hemocyanin-derived phenoloxidase, inducing conformational changes that permit phenolic substrate access to the active site. Here, we observed extracellular hemocyanin-derived phenoloxidase activity levels increase in the presence of apoptotic amebocytes. Enzyme activity induced by phosphatidylserine or apoptotic amebocytes was reduced completely upon incubation with the phosphatidylserine binding protein, annexin V. We propose that phosphatidylserine redistributed to the outer plasma membrane of amebocytes undergoing phagocytosis-induced apoptosis could interact with hemocyanin, thus facilitating its conversion into a phenoloxidase-like enzyme, during immune challenge.
Biological and Environmental Sciences, School of Natural Sciences, University of Stirling, Stirling, FK9 4LA, Scotland, UK. c.j.coates@stir.ac.uk.
Christopher J
CJ
Coates
Biological and Environmental Sciences, School of Natural Sciences, University of Stirling, Stirling, FK9 4LA, Scotland, UK.
Tim
T
Whalley
Biological and Environmental Sciences, School of Natural Sciences, University of Stirling, Stirling, FK9 4LA, Scotland, UK.
Michael
M
Wyman
Biological and Environmental Sciences, School of Natural Sciences, University of Stirling, Stirling, FK9 4LA, Scotland, UK. jacqueline.nairn@stir.ac.uk.
Jacqueline
J
Nairn
D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis; D052982:Beauveria / Q000472:pathogenicity / Q000502:physiology; D020169:Caspases / Q000235:genetics / Q000378:metabolism; D053938:DNA Fragmentation; D004789:Enzyme Activation; D005786:Gene Expression Regulation; D006433:Hemocyanins / Q000235:genetics / Q000378:metabolism*; D006737:Horseshoe Crabs / Q000201:enzymology / Q000235:genetics* / Q000276:immunology / Q000382:microbiology; D054884:Host-Pathogen Interactions; D014442:Monophenol Monooxygenase / Q000235:genetics / Q000378:metabolism*; D010586:Phagocytes / Q000276:immunology / Q000382:microbiology*; D010587:Phagocytosis; D010718:Phosphatidylserines / Q000378:metabolism; D013172:Spores, Fungal / Q000472:pathogenicity / Q000502:physiology
D017304:Annexin A5; D010718:Phosphatidylserines; D006433:Hemocyanins; D014442:Monophenol Monooxygenase; D020169:Caspases