Serpin-1 splicing isoform J inhibits the proSpätzle-activating proteinase HP8 to regulate expression of antimicrobial hemolymph proteins in Manduca sexta.
Serpin-1 splicing isoform J inhibits the proSpätzle-activating proteinase HP8 to regulate expression of antimicrobial hemolymph proteins in Manduca sexta.
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DOI：
10.1016/j.dci.2010.09.004
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发表时间：
2011-01
影响因子：
2.9
通讯作者：
Kanost, Michael R.
中科院分区：
文献类型：
--
作者：
An, Chunju;Ragan, Emily J.;Kanost, Michael R.
昆虫的先天免疫系统包括Toll通路，它是由细胞外丝氨酸蛋白酶级联介导的。在烟草角虫（Manduca sexta）中，血淋巴蛋白酶8 （HP8）通过切割proSpätzle促进抗菌蛋白的合成，proSpätzle被认为是sexta Toll的配体。HP8在血淋巴中与M. sexta serin -1形成复合体，该复合体具有多种可选择的剪接异构体。为了研究HP8的调控及其对proSpätzle的加工，我们对重组HP8与serpin-1亚型J （serpin-1J）的相互作用进行了表征。重组蛇蛋白1j在体外与HP8形成sds稳定的复合物。serin - 1j与HP8的结合速率常数为1.3×104 M-1s-1，抑制化学计量值为5.4。serin - 1j抑制了HP8对proSpätzle的裂解。注射丝氨酸- 1j后，血淋巴中细菌诱导的抗菌活性降低，脂肪体中丝氨酸、黏附素和血红素mRNA表达降低。综上所述，这些结果表明，serpin-1J可以抑制HP8，从而调节活性Spätzle的浓度，从而调节m.s sexta的Toll通路反应。
The innate immune system of insects include the Toll pathway, which is mediated by an extracellular serine proteinase cascade. In the tobacco hornworm, Manduca sexta, hemolymph proteinase 8 (HP8) promotes the synthesis of antimicrobial proteins by cleaving proSpätzle, the putative ligand of M. sexta Toll. HP8 has been observed to form a complex in hemolymph with M. sexta serpin-1, which has multiple alternative splicing isoforms. To investigate the regulation of HP8 and its processing of proSpätzle, we characterized the interaction of recombinant HP8 with serpin-1 isoform J (serpin-1J). Recombinant serpin-1J formed an SDS-stable complex with HP8 in vitro. The association rate constant of serpin-1J and HP8 was 1.3×104 M-1s-1, with a stoichiometry of inhibition of 5.4. Serpin-1J inhibited the cleavage of proSpätzle by HP8. Injection of serpin-1J into M. sexta larvae resulted in decreased bacteria-induced antimicrobial activity in hemolymph and reduced expression of cecropin, attacin and hemolin mRNA in fat body. Altogether, these results suggest that serpin-1J functions to inhibit HP8 and thereby modulates the concentration of active Spätzle to regulate the Toll pathway response in M. sexta.
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DOI：
10.1073/pnas.0610924104
发表时间：
2007-04-17
影响因子：
11.1
作者：
Park, Ji-Won;Kim, Chan-Hee;Lee, Bok-Luel
通讯作者：
Lee, Bok-Luel
DOI：
10.1073/pnas.0903134106
发表时间：
2009-07-21
影响因子：
11.1
作者：
Ahmad, S. Tariq;Sweeney, Sean T.;Gao, Fen-Biao
通讯作者：
Gao, Fen-Biao
影响因子：
56.9
作者：
Levashina, EA;Langley, E;Reichhart, JM
通讯作者：
Reichhart, JM
影响因子：
4.8
作者：
Jiang, HB;Wang, Y;Kanost, MR
通讯作者：
Kanost, MR
影响因子：
3.8
作者：
KANOST, MR;PRASAD, SV;WILLOTT, E
通讯作者：
WILLOTT, E