大黄粉虫幼虫体内黑色素合成相关蛋白的鉴定

目的鉴定大黄粉虫幼虫体内与酚氧化酶(PO)诱导的黑色素合成相关的蛋白(MAPs)。方法体外诱导大黄粉虫幼虫血淋巴中黑色素合成反应,对上清液中的蛋白质进行SDS PAGE分析,比较黑色素合成前后、合成过程中及在苯硫脲存在下蛋白质的变化,对黑色素合成过程中消失的蛋白进行氨基端氨基酸序列分析。结果6条蛋白带(MAP 1~6)呈时间依赖性地从黑色素合成反应的上清液中消失,苯硫脲可阻断此反应而取消蛋白的消失;氨基端氨基酸序列分析表明MAP 1,2为已知的大黄粉虫卵黄素样蛋白,MAP 3,4,5为未知蛋白,MAP 6为已知的大黄粉虫干燥蛋白。结论大黄粉虫幼虫血淋巴中至少5种蛋白与PO诱导的黑色素合成反应特异性相关。

Identification of melanization-associated proteins in Tenebrio molitor larvae

Objective To identify the proteins associated with PO-induced melanin synthesis(MAPs) in Tenebrio molitor larvae.Methods The melanin synthesis of G-100 solution was induced by β-1,3-D-glucan and Ca~(2+) in the presence of dopamine in vitro.The proteins in the supernatant of the reaction mixture were analyzed by 12% SDS-PAGE electrophoresis.The amino-terminal amino acid sequences of the proteins disappeared during the PO-induced melanin synthesis were determined.Results Six proteins disappeared from the supernatant of the melanization reaction mixture in a time-dependent manner.In the presence of PTU,an inhibitor of PO activity,the six bands did not disappear.The amino-terminal sequence analysis showed that both MAP-1 and 2 were the vitellogenin-like protein of T.molitor.The MAP-3,4 and 5 were unknown proteins.The MAP-6 was the desiccation protein of T.molitor.Conclusions At least 5 proteins are specially associated with PO-induced melanin synthesis in T.molitor larvae.