Characterisation of proteases involved in egg hatching of the sheep blowfly, Lucilia cuprina.
A number of proteases were identified in the egg shell washings (ESW) collected during the egg hatching of Lucilia cuprina (sheep blowfly). Characterization of these proteases indicated a pH optima in a similar pH range that was optimal for L. cuprina egg hatching. Mechanistic characterization of these proteases indicated that they were predominantly of the serine class. Several protease inhibitors were tested for their ability to inhibit L. cuprina egg hatching in vitro. Egg hatching was significantly (P<0.05) inhibited by PMSF (61%), 1,10-Phenanthroline (42%) and Pepstatin (29%). The inhibition of egg hatching by PMSF showed a strong concentration dependence, with its effects ranging from inhibition at high concentrations to enhancement of egg hatching at low concentrations. Addition of ESW to unhatched eggs, significantly (P<0.05) enhanced their rate of hatching above untreated control eggs. This enhancement of egg hatching was significantly (P<0.05) reversed by the protease inhibitors Elastatinal (40%), 1,10-Phenanthroline (40%) and PMSF (38%). These studies indicate a role for serine and/or metallo-proteases in facilitating L. cuprina egg hatch.
Young
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AR
Centre for Animal Biotechnology, School of Veterinary Science, The University of Melbourne, Victoria 3052, Parkville, Australia.
Mancuso
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Meeusen
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EN
Bowles
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VM
D000818:Animals; D004175:Diptera / Q000187:drug effects / Q000201:enzymology / Q000502:physiology*; D010450:Endopeptidases / Q000145:classification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D011480:Protease Inhibitors / Q000494:pharmacology
D011480:Protease Inhibitors; D010450:Endopeptidases