• Title/Summary/Keyword: Apolipophorin-III

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Apolipophorin-III uptake by the adult testes in the wax moth, Galleria mellonella (꿀벌부채명나방 성충 정소에 의한 아포리포포린-III의 흡수)

  • Yun, Hwa-Kyung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.17 no.10
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    • pp.199-203
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    • 2016

  • Apolipophorin-III (apoLp-III) was isolated and purified from the last larval hemolymph of Galleria mellonella by KBr gradient ultracentrifugation and gel chromatography (Sephadex G-100). After KBr gradient ultracentrifugation, the lipophorin-free fractions were used as the samples for gel chromatography. The purity of the finally purified apoLp-III was confirmed by SDS-PAGE after gel chromatography. In this study, we found that apoLp-III is taken up into the adult testes in Galleria mellonella. The testes were dissected from day-1 or -2 adults in cold Ringer's solution and used for tissue culture. The protein moiety of apoLp-III was labeled with FITC dissolved in dimethyl sulfoxide (DMSO) at room temperature under conditions of continuous stirring for 1 h. The FITC-labeled apoLp-III was purified with a Sephadex G-25 PD-10 column. The tissues of the adult testes were incubated at room temperature for 30 min with fluorescein isothiocyanate (FITC)-labeled apoLp-III. Fluorescein microscopy and sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) revealed that the adult testis tissues internalize the FITC-labeled apoLp-III. The results showed that apoLp-III is taken up by the adult testes.

  • https://doi.org/10.5762/KAIS.2016.17.10.199 인용 PDF KSCI

Degradation of Insect Humoral Immune Proteins by the Proteases Secreted from Enterococcus faecalis

  • Park, Shin-Yong;Kim, Koung-Mi;Kim, Ik-Soo;Lee, Sang-Dae;Lee, In-Hee
    • International Journal of Industrial Entomology and Biomaterials
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    • v.13 no.1
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    • pp.37-43
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    • 2006

  • Enterococcus faecalis was isolated from the body fluid of dead Galleria mellonella larvae. Upon injection of E. faecalis into the hemocoel of G. mellonella, the bacteria destroyed parts of humoral defense systems in the hemolymph. In a test for the proteolytic activity of E. faecalis CS, it was confirmed that the enzyme degraded three well-known a-helical antimicrobial peptides, cecropin A, melittin and halocidin, and abolished their activities. We also determined putative cleavage sites on the primary sequences of three peptides through purification and mass analysis of peptide fragments digested by E. faecalis CS. Furthermore it was found that apolipophorin-III, recently known as a critical recognition protein for invading microbes in the hemolymph of G. mellonella, was also degraded by E. faecalis CS. Taken together, the present work shows that the protease in secretions from E. faecalis destroyed two critical humoral immune factors in the hemolymph of G. mellonella larvae. In addition, this paper demonstrates that the relationship between the host insect and the pathogenic bacteria might provide a valuable model system to study the enterococcal virulence mechanism, which may be relevant to mammalian pathogenesis.

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