• Title/Summary/Keyword: Hemolymph protein

Search Result 92, Processing Time 0.044 seconds

Purification and characterization of a 1,3-β-D-glucan recognition protein from Antheraea pernyi larve that is regulated after a specific immune challenge

  • Youlei, Ma;Jinghai, Zhang;Yuntao, Zhang;Jiaoshu, Lin;Tianyi, Wang;Chunfu, Wu;Rong, Zhang
    • BMB Reports
    • /
    • v.46 no.5
    • /
    • pp.264-269
    • /
    • 2013
  • Pattern recognition receptors are known to participate in the activation of Prophenoloxidase system. In this study, a 1,3-${\beta}$-D-glucan recognition protein was detected for the first time in Antheraea pernyi larvae (Ap-${\beta}GRP$). Ap-${\beta}GRP$ was purified to 99.9% homogeneity from the hemolymph using traditional chromatographic methods. Ap-${\beta}GRP$ specifically bind 1,3-${\beta}$-D-glucan and yeast, but not E. coli or M. luteus. The 1,3-${\beta}$-D-glucan dependent phenoloxidase (PO) activity of the hemolymph inhibited by anti-Ap-${\beta}GRP$ antibody could be recovered by addition of purified Ap-${\beta}GRP$. These results demonstrate that Ap-${\beta}GRP$ acts as a biosensor of 1,3-${\beta}$-Dglucan to trigger the Prophenoloxidase system. A trace mount of 1,3-${\beta}$-D-glucan or Ap-${\beta}GRP$ alone was unable to trigger the proPO system, but they both did. Ap-${\beta}GRP$ was specifically degraded following the activation of proPO with 1,3-${\beta}$-Dglucan. These results indicate the variation in the amount of Ap-${\beta}GRP$ after specific immune challenge in A. pernyi hemolymph is an important regulation mechanism to immune response.

Electrophoretic Patterns of Hemolymph Protein of Healthy and Nuclear-Polyhedrosis Virus Infected Silkworm, Bombyx mori. (건강잠 및 핵다각체 이병잠의 체액단백질의 전기영동상)

  • 김낙정;강석권;김정일
    • Journal of Sericultural and Entomological Science
    • /
    • v.13 no.2
    • /
    • pp.141-143
    • /
    • 1971
  • By means of thin-layer electrophoresis in agarose gel, hemolymph protein of healthy silkworm larvae and of the nuclear polygedrosis virus infected larvae were studied. 1. In the 4th instar, 4 fractions moving toward anode were separated. Dye-binding Capacity of the fraction was increased according to the stage. 2. After 5th day in the 5th instar, 7 fractions moving toward anode were separated, and one fraction toward cathode was separated. 3. On the first day in the 5th instar, 5 fractions were separated, and on the 4th day of the same instar 5 fractions were separated. 4. As for the hemolymph protein fractions of the polyhedrosis virus infected larvae, on the 6th and 7th day, three fractions(D.E.F) were inclined to increase, whereas on the 8th day 4 fractions(A.B.D.E) were disappeared but F fraction was inclined to decrease.

  • PDF

One-step Separation of 30K Protein from the Silkworm Hemolymph by Anion-exchange Chromatography and Its Effect on the Proliferation of Human Cells (음이온교환 크로마토그래피를 이용한 누에체액 유래 30K 단백질의 정제와 정제된 단백질이 인간세포 배양 증식에 미치는 영향)

  • Shin Hyun-Chong;Joung Chan-Hi;Choi Yong-Soo;Lim Sang-Min;Han Kyuboem;Koo Yoon-Mo;Park Tai Hyun;Kim Dong-Il
    • KSBB Journal
    • /
    • v.20 no.3
    • /
    • pp.233-237
    • /
    • 2005
  • In order to investigate the feasibility of 30K protein from silkworm (Bombyx mori) hemolymph (SH) on the proliferation of human cells, a simple separating procedure by anion-exchange chromatography system with Q-Sepharose fast flow gel was established. The 30K protein was eluted with an optimized condition of 0.16 M sodium chloride in 20 mM tris buffer (pH 9.0). The separated 30K protein at three concentrations of 0.04, 0.12, and 0.4 mg/ml was added to the culture medium with various human cells, such as chondrocytes, periosteum-derived cells, and MRC-5 cells, and their growth rates were measured. The cell growth rate at protein concentration of 0.4 mg/ml was always higher than that without 30K protein in all human cells tested, suggesting that the 30K protein has positive effect on the increase of the life span of human cells.

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
    • /
    • v.13 no.1
    • /
    • pp.37-43
    • /
    • 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.

Exosome isolation from hemolymph of white-spotted flower chafer, Protaetia brevitarsis (Kolbe) (Coleoptera: Scarabaeidae).

  • Lee, Seokhyun;Kwon, Kisang;Song, Myung-Ha;Park, Kwan-ho;Kwon, O-Yu;Choi, Ji-young
    • International Journal of Industrial Entomology and Biomaterials
    • /
    • v.33 no.2
    • /
    • pp.85-91
    • /
    • 2016
  • Exosomes are homogenous vesicles of 40-100 nm diameter produced endogenously. Exosomes are generated by inward budding into multi-vesicular bodies (MVB) and then released to extracellular space. Exosomes contain various nucleic acid and protein cargoes from their cells of origin and this endosomal cellular molecules are used for intracellular communication and for both promotion and suppression of immune responses. Recently, they are also considered as delivery vehicle for therapeutic proteins due to their characteristics of stability in body fluids and ability for target uptake. Also, they show less immune reactivity because the isolated exosome harboring therapeutic proteins can be from the same host. White-spotted flower chafer, Protaetia brevitarsis is one of the major insect commercially reared in Korea. There are bacterial and fungal pathogens causing diseases in the beetle, and these diseases incur economic loss to the larva-rearing farms. Due to their endosomal cargoes, exosomes are good candidates in use of disease diagnosis. In this study, we isolated insect exosome from the hemolymph of P. brevitarsis, and verified it by analysis of the exosome-specific surface proteins and RNA.