Scholarlink

<p>A new protein with immune properties was found in <i>Galleria mellonella</i> hemolymph. The so-far putative serine protease inhibitor dipetalogastin-like (GmSPID) was found in one fraction obtained after separation of hemolymph by RP-HPLC. Its amount depended on the immune status of the insect: it significantly increased after oral (10^3 CFU) and intrahemocelic (10 and 50 CFU) infection with entomopathogenic bacteria <i>Pseudomonas entomophila</i>. This was accompanied by up-regulation of the respective gene in the fat body of infected larvae. GmSPID was purified to homogeneity and characterised as a protein with immune properties. Among the three proteases tested, i.e. trypsin, elastase, and thermolysin, the strongest inhibition was observed toward trypsin. No inhibition toward the metalloproteinase thermolysin was detected, confirming that GmSPID is an inhibitor of serine proteases. Additionally, GmSPID was shown to have antimicrobial properties. At the concentration of 7 µM and 15 µM, it acted against <i>Pseudomonas entomophila</i>, <i>Pseudomonas aeruginosa</i>, <i>Bacillus thuringiensis</i>, <i>Escherichia coli</i>, and <i>Candida albicans</i> but not against <i>Staphylococcus aureus</i>. Moreover, with the use of atomic force, scanning, and transmission electron microscopy techniques, we present the effect of the GmSPID protein on the surface properties, shape, and ultrastructure of <i>P. entomophila</i> cells. The protein caused modest perforation of the cellular membrane, contributing to loss of its integrity. The mode of the GmSPID protein action as an antimicrobial compound and its role in <i>G. mellonella</i> immunity are discussed.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Serine protease inhibitor dipetalogastin-like from Galleria mellonella is involved in insect immunity

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2025年7月6日

查看全文
  • Jakub Kordaczuk,
  • Michał Sułek,
  • Paweł Mak,
  • Bożena Pawlikowska-Pawlęga,
  • Iwona Wojda

摘要

A new protein with immune properties was found in Galleria mellonella hemolymph. The so-far putative serine protease inhibitor dipetalogastin-like (GmSPID) was found in one fraction obtained after separation of hemolymph by RP-HPLC. Its amount depended on the immune status of the insect: it significantly increased after oral (10^3 CFU) and intrahemocelic (10 and 50 CFU) infection with entomopathogenic bacteria Pseudomonas entomophila. This was accompanied by up-regulation of the respective gene in the fat body of infected larvae. GmSPID was purified to homogeneity and characterised as a protein with immune properties. Among the three proteases tested, i.e. trypsin, elastase, and thermolysin, the strongest inhibition was observed toward trypsin. No inhibition toward the metalloproteinase thermolysin was detected, confirming that GmSPID is an inhibitor of serine proteases. Additionally, GmSPID was shown to have antimicrobial properties. At the concentration of 7 µM and 15 µM, it acted against Pseudomonas entomophila, Pseudomonas aeruginosa, Bacillus thuringiensis, Escherichia coli, and Candida albicans but not against Staphylococcus aureus. Moreover, with the use of atomic force, scanning, and transmission electron microscopy techniques, we present the effect of the GmSPID protein on the surface properties, shape, and ultrastructure of P. entomophila cells. The protein caused modest perforation of the cellular membrane, contributing to loss of its integrity. The mode of the GmSPID protein action as an antimicrobial compound and its role in G. mellonella immunity are discussed.