Involvement of peroxidase in chorion hardening in Aedes aegypti
Involvement of peroxidase in chorion hardening in Aedes aegypti
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DOI：
10.1016/0965-1748(95)00099-2
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发表时间：
1996-03-01
影响因子：
3.8
通讯作者：
Christensen, BM
中科院分区：
文献类型：
--
作者：
Li, JY;Hodgeman, BA;Christensen, BM
在埃及伊蚊卵巢中，含有发育中的卵，在吸血后24小时可检测到过氧化物酶活性，并且在血餐后36-48小时达到峰值过氧化物酶活性。过氧化物酶与成熟卵子中的绒毛膜层相关，并且通过用SDS/尿素处理分离的绒毛膜部分，大部分酶从绒毛膜层释放。使用具有托酚酮/H2 O2或多巴染色的SDS-PAGE分析SDS/尿素溶解的绒毛膜蛋白证实了在释放的绒毛膜蛋白中存在过氧化物酶和酚氧化酶。绒毛膜过氧化物酶的分子量为约61，000 Da，如通过SDS-PAGE分析所测定的。溶解的绒毛膜蛋白与酪氨酸和H2 O2孵育产生二酪氨酸，结果表明，绒毛膜过氧化物酶通过结构蛋白上的酪氨酸残基催化二酪氨酸的形成，参与蚊卵绒毛膜的硬化。埃及卵绒毛膜包括通过双酪氨酸形成的过氧化物酶介导的绒毛膜蛋白交联和酚氧化酶催化的绒毛膜黑化。
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.