Abstract:
After separation on gel zymography, Drosophila melanogaster hemolymph displays gelatinase and caseinase bands of varying sizes, ranging from over 140 to 25 kDa. Qualitative and quantitative variations in these bands were observed during larval development and between different D. melanogaster strains and Drosophila species. The activities of these Drosophila hemolymph gelatinase and caseinase were strongly inhibited by serine protease inhibitors, but not by EDTA. Mass spectrometry identified over 60 serine proteases (SPs) in gel bands corresponding to the major D. melanogaster gelatinases and caseinases, but no matrix metalloproteinases (MMPs) were found. The most abundant proteases were tequila and members of the Jonah and trypsin families. However, the gelatinase bands did not show any change in the tequila null mutant. Additionally, no clear changes could be observed in D. melanogaster gel bands 24 h after injection of bacterial lipopolysaccharides (LPS) or after oviposition by Leptopilina boulardi endoparasitoid wasps. It can be concluded that the primary gelatinases and caseinases in Drosophila larval hemolymph are serine proteases (SPs) rather than matrix metalloproteinases (MMPs). Furthermore, the gelatinase pattern remains relatively stable even after short-term exposure to pathogenic challenges.
摘要:
在凝胶酶谱上分离后,果蝇血淋巴显示不同大小的明胶酶和酪蛋白酶带,范围从140到25kDa。在幼虫发育过程中以及在不同的D.melanogaster菌株和果蝇物种之间观察到这些条带的定性和定量变化。这些果蝇血淋巴明胶酶和酪蛋白酶的活性被丝氨酸蛋白酶抑制剂强烈抑制,但不是EDTA。质谱鉴定出超过60个丝氨酸蛋白酶(SP)在凝胶带中对应于主要的D.melanogaster明胶酶和酪蛋白酶,但没有发现基质金属蛋白酶(MMPs)。最丰富的蛋白酶是龙舌兰酒和乔纳和胰蛋白酶家族的成员。然而,明胶酶带在龙舌兰酒无效突变体中没有显示任何变化。此外,在注射细菌脂多糖(LPS)后24小时或Leptopilinaboulardi内寄生虫类黄蜂产卵后,未观察到D.melanogaster凝胶带的明显变化。可以得出结论,果蝇幼虫血淋巴中的主要明胶酶和酪蛋白蛋白酶是丝氨酸蛋白酶(SP),而不是基质金属蛋白酶(MMPs)。此外,即使在短期暴露于致病性挑战后,明胶酶模式仍保持相对稳定。