A positive feedback mechanism in the Manduca sexta prophenoloxidase activation system.
In Manduca sexta, pathogen recognition triggers a branched serine proteinase cascade which generates active phenoloxidase (PO) in the presence of a proPO-activating proteinase (PAP) and two noncatalytic serine proteinase homologs (SPHs). PO then catalyzes the production of reactive compounds for microbe killing, wound healing, and melanin formation. In this study, we discovered that a minute amount of PAP1 (a final component of the proteinase pathway) caused a remarkable increase in PO activity in plasma from naïve larvae, which was significantly higher than that from the same amounts of PAP1, proPO and SPHs incubated in vitro. The enhanced proPO activation concurred with the proteolytic activation of HP6, HP8, PAP1, SPH1, SPH2 and PO precursors. PAP1 cleaved proSPH2 to yield bands with mobility identical to SPH2 generated in vivo. PAP1 partially hydrolyzed proHP6 and proHP8 at a bond amino-terminal to the one cut in the PAP1-added plasma. PAP1 did not directly activate proPAP1. These results suggest that a self-reinforcing mechanism is built into the proPO activation system and other plasma proteins are required for cleaving proHP6 and proHP8 at the correct site to strengthen the defense response, perhaps in the early stage of the pathway activation.
Wang
Yang
Y
Department of Entomology and Plant Pathology, Oklahoma State University, Stillwater, OK 74078, USA.
Jiang
Haobo
H
D000818:Animals; D004156:Catechol Oxidase / Q000378:metabolism*; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000378:metabolism*; D025461:Feedback, Physiological; D006458:Hemolymph / Q000201:enzymology*; D015151:Immunoblotting; D018402:Manduca / Q000201:enzymology* / Q000276:immunology; D000073718:Pancreatitis-Associated Proteins; D010447:Peptide Hydrolases / Q000276:immunology / Q000378:metabolism; D012697:Serine Endopeptidases / Q000378:metabolism*
D004792:Enzyme Precursors; D000073718:Pancreatitis-Associated Proteins; C000612141:REG3A protein, human; C038451:pro-phenoloxidase; D004156:Catechol Oxidase; D010447:Peptide Hydrolases; D012697:Serine Endopeptidases; C115269:pro-phenol oxidase activating proteinase