Nauen R, Sorge D, Sterner A, Borovsky D
Bayer AG, Agrochemicals Division, Research Insecticides, Leverkusen, Germany.
Arch Insect Biochem Physiol. 2001 Aug;47(4):169-80. doi: 10.1002/arch.1049.
Proteolytic enzyme biosynthesis in the midgut of the 4th instar larva of Heliothis virescens is cyclical. Protease activity increases immediately after the molt from the 3rd to the 4th instar larvae and declines just before the molt into the 5th instar. Characterization of the midgut proteases using soybean tryspin inhibitor (SBTI) Bowman Birk Inhibitor (BBI) 4-(2-aminoethyl)benzensulfonylfluoride (AEBSF) and N-tosyl-L-phenylalanine chloromethylketone (TPCK) indicate that protease activity is mostly trypsinlike (80%) with a small amount of chymotrypsinlike activity (20%). Injections of late 3rd and 4th instar larval hemolymph into H. virescens larvae inhibited tryspin biosynthesis in the larval midgut. Similar results were obtained when highly purified 4th instar larval hemolymph that crossreacted with Aea-TMOF antisurm using ELISA was injected into 2nd instar larvae. Injections of Aea-TMOF and its analogues into 2nd instar, and Aea-TMOF alone into 4th instar larvae stopped trypsin biosynthesis 24 and 48 h after the injections, respectively. Injections of 4th instar H. virescens larval hemolymph into female Aedes aegypti that took a blood meal stopped trypsin biosynthesis and egg development. These results show that the biosynthesis of trypsin-like enzymes in the midgut of a lepidoptera is modulated with a hemolymph circulating TMOF-like factor that is closely related to Aea-TMOF. Arch.
棉铃虫四龄幼虫中肠蛋白水解酶的生物合成是周期性的。蛋白酶活性在从三龄幼虫蜕皮至四龄幼虫后立即增加,并在蜕皮至五龄幼虫前下降。使用大豆胰蛋白酶抑制剂(SBTI)、鲍曼-伯克抑制剂(BBI)、4-(2-氨基乙基)苯磺酰氟(AEBSF)和N-对甲苯磺酰-L-苯丙氨酸氯甲基酮(TPCK)对中肠蛋白酶进行表征表明,蛋白酶活性主要为类胰蛋白酶(80%),伴有少量类糜蛋白酶活性(20%)。将三龄晚期和四龄幼虫的血淋巴注射到棉铃虫幼虫中,可抑制幼虫中肠胰蛋白酶的生物合成。当使用酶联免疫吸附测定(ELISA)与埃及伊蚊TMOF(Aea-TMOF)抗血清发生交叉反应的高度纯化的四龄幼虫血淋巴注射到二龄幼虫中时,也获得了类似的结果。将Aea-TMOF及其类似物注射到二龄幼虫中,以及将Aea-TMOF单独注射到四龄幼虫中,分别在注射后24小时和48小时停止了胰蛋白酶的生物合成。将棉铃虫四龄幼虫的血淋巴注射到已取食的雌性埃及伊蚊中,可停止胰蛋白酶的生物合成并抑制卵子发育。这些结果表明,鳞翅目昆虫中肠类胰蛋白酶的生物合成受到一种与血淋巴循环的TMOF样因子的调节,该因子与Aea-TMOF密切相关。《Archives》