Nuss Andrew B, Gulia-Nuss Monika
Department of Biochemistry and Molecular Biology, Veterinary, and Rangeland Sciences, University of Nevada, Reno, Nevada 89557, USA.
Department of Agriculture, Veterinary, and Rangeland Sciences, University of Nevada, Reno, Nevada 89557, USA.
Cold Spring Harb Protoc. 2023 Apr 3;2023(4):pdb.top107656. doi: 10.1101/pdb.top107656.
When a female mosquito takes a blood meal, proteolytic activity surges in the midgut. Trypsin-like serine proteases are the major endoproteolytic enzyme induced by feeding in mosquitoes. The mosquito midgut lacks trypsin activity before the blood meal, but in most anautogenous mosquitoes, trypsin activity increases continuously up to 30 h after feeding and subsequently returns to baseline levels by 60 h. Trypsin activity in mosquitoes is restricted entirely to the posterior midgut lumen, where blood is stored and digested. Trypsin enzyme activity can be quantitatively measured using the artificial α-benzoyl--arginine 4-nitroanilide hydrochloride substrate, a method described in our associated protocol.
当雌性蚊子吸食血液时,中肠中的蛋白水解活性激增。类胰蛋白酶丝氨酸蛋白酶是蚊子进食后诱导产生的主要内切蛋白酶。在吸食血液之前,蚊子的中肠缺乏胰蛋白酶活性,但在大多数非自育性蚊子中,胰蛋白酶活性在进食后持续增加,直至30小时,随后在60小时恢复到基线水平。蚊子体内的胰蛋白酶活性完全局限于中肠后段管腔,血液在那里储存和消化。胰蛋白酶的酶活性可以使用人工α-苯甲酰-L-精氨酸4-硝基苯胺盐酸盐底物进行定量测量,这是我们相关实验方案中描述的一种方法。