Involvement of peroxidase in chorion hardening in Aedes aegypti.
Peroxidase activity is detectable in Aedes aegypti ovaries, containing developing eggs, at 24 h following blood feeding, and peak peroxidase activity is reached at 36-48 h after the blood-meal. Peroxidase is associated with the chorion layer in mature eggs and the majority of the enzyme is released from the chorion layer by treating the isolated chorion fraction with SDS/urea. Analysis of the SDS/urea solubilized chorion proteins using SDS-PAGE with tropolone/H2O2 or dopa staining verified the presence of both peroxidase and phenol oxidase in the released chorion proteins. The molecular weight of chorion peroxidase is about 61,000 Da as determined by SDS-PAGE analysis. Incubation of the solubilized chorion proteins with tyrosine and H2O2 produces dityrosine, and hyrolysis of hardened egg chorion results in the detection of dityrosine and trityrosine in the chorion hydrolysate. Data suggest that chorion peroxidase is involved in the hardening of the mosquito egg chorion by catalyzing the formation of ditryrosine through tyrosine residues on structural proteins. The overall hardening of the A. aegypti egg chorion includes both peroxidase-mediated chorion protein crosslinking through dityrosine formation and phenol oxidase-catalyzed chorion melanization.
Department of Animal Health and Biomedical Sciences, University of Wisconsin-Madison 53706, U.S.A.
J
J
Li
B A
BA
Hodgeman
B M
BM
Christensen
D000330:Aedes / Q000201:enzymology* / Q000254:growth & development; D000818:Animals; D002823:Chorion / Q000378:metabolism*; D004527:Egg Proteins / Q000378:metabolism*; D005260:Female; D006861:Hydrogen Peroxide / Q000494:pharmacology; D010063:Ovum / Q000201:enzymology* / Q000254:growth & development; D009195:Peroxidase / Q000302:isolation & purification / Q000378:metabolism*; D013997:Time Factors; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism
D004527:Egg Proteins; C015753:chorion proteins; D014443:Tyrosine; D006861:Hydrogen Peroxide; C007543:dityrosine; D009195:Peroxidase