Kazal-type serine proteinase inhibitors in the midgut of Phlebotomus papatasi.
Kazal-type serine proteinase inhibitors in the midgut of Phlebotomus papatasi.
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DOI：
10.1590/0074-0276108062013001
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发表时间：
2013-09
影响因子：
2.8
通讯作者：
Ramalho-Ortigão M
中科院分区：
文献类型：
--
作者：
Sigle LT;Ramalho-Ortigão M
沙蝇是利什曼原虫、细菌和病毒的重要病媒。通过对主要利什曼原虫载体--罂粟白头翁的中肠转录组的研究，鉴定出两个非经典的Kazal型丝氨酸蛋白酶抑制物(PpKzl1和PpKzl2)。表达谱分析表明，PpKzl1和PpKzl2转录本均受中肠采血的调节，在雄虫、幼虫和蛹中也有表达。我们在哺乳动物表达系统(CHO-S游离细胞)中表达了重组PpKzl2，并将其用于体外研究丝氨酸蛋白酶的抑制作用。重组PpKzl2对α-糜蛋白酶的抑制作用为9.4%，对α-凝血酶和胰酶的抑制作用分别为33.5%和63.9%，提示天然PpKzl2是一种具有活性的丝氨酸蛋白酶抑制物，可能参与调节中肠消化酶。利什曼原虫的早期阶段很容易被沙蝇中肠的消化酶杀死。因此，确定丝氨酸蛋白酶抑制剂的特征可能为预防利什曼原虫的传播提供新的靶点和策略。
Sandflies (Diptera: Psychodidae) are important disease vectors of parasites of the genus Leishmania, as well as bacteria and viruses. Following studies of the midgut transcriptome of Phlebotomus papatasi, the principal vector of Leishmania major, two non-classical Kazal-type serine proteinase inhibitors were identified (PpKzl1 and PpKzl2). Analyses of expression profiles indicated that PpKzl1 and PpKzl2 transcripts are both regulated by blood-feeding in the midgut of P. papatasi and are also expressed in males, larva and pupa. We expressed a recombinant PpKzl2 in a mammalian expression system (CHO-S free style cells) that was applied to in vitro studies to assess serine proteinase inhibition. Recombinant PpKzl2 inhibited α-chymotrypsin to 9.4% residual activity and also inhibited α-thrombin and trypsin to 33.5% and 63.9% residual activity, suggesting that native PpKzl2 is an active serine proteinase inhibitor and likely involved in regulating digestive enzymes in the midgut. Early stages of Leishmania are susceptible to killing by digestive proteinases in the sandfly midgut. Thus, characterising serine proteinase inhibitors may provide new targets and strategies to prevent transmission of Leishmania.
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DOI：
10.1046/j.1432-1327.1999.00895.x
发表时间：
1999-12-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
Mende, K;Petoukhova, O;Nowak, G
通讯作者：
Nowak, G
影响因子：
3.8
作者：
Coutinho-Abreu IV;Sharma NK;Robles-Murguia M;Ramalho-Ortigao M
通讯作者：
Ramalho-Ortigao M
影响因子：
4.4
作者：
Jochim RC;Teixeira CR;Laughinghouse A;Mu J;Oliveira F;Gomes RB;Elnaiem DE;Valenzuela JG
通讯作者：
Valenzuela JG
影响因子：
4
作者：
Araujo, R. N.;Campos, I. T. N.;Pereira, M. H.
通讯作者：
Pereira, M. H.
DOI：
10.1046/j.1432-1327.2001.02084.x
发表时间：
2001-04-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
Nirmala, X;Kodrik, D;Sehnal, F
通讯作者：
Sehnal, F