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Glutathione-S-transferase alpha-4 in Hippocampus abdominalis (big-belly seahorse): Molecular characterization, antioxidant properties, and its potent immune response

첫 페이지 보기
  • 발행기관
    제주대학교 해양과학연구소 바로가기
  • 간행물
    해양과학연구소 연구논문집 바로가기
  • 통권
    제45권 (2021.12)바로가기
  • 페이지
    pp.41-51
  • 저자
    Kishanthini Nadarajapillai, D.S. Liyanage, Sarithaa Sellaththurai, Taehyug Jeong, Sukkyoung Lee, Jehee Lee
  • 언어
    영어(ENG)
  • URL
    https://www.earticle.net/Article/A406934

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원문정보

초록

영어
Glutathione-S-transferase (GST) is a key enzyme in the phase-II detoxification process and is a biomarker of oxidative stress. In this study, we analyzed the molecular, biochemical, and antioxidant properties of GST alpha-4 from Hippocampus abdominalis (HaGSTA-4). Also, the spatial and temporal expression of HaGSTA-4 upon immune challenge with abiotic and biotic stimulants were evaluated. The HaGSTA-4 ORF encodes 223 amino acids with a molecular weight of 25.7 kDa, and an estimated isoelectric point (pI) of 8.47. It consists of the GST_C superfamily and thioredoxin-like superfamily domain. The phylogenetic tree revealed that HaGSTA-4 is evolutionarily conserved with its GST alpha class counterparts. From pairwise alignment, the highest values of identity (78.5%) and similarity (85.7%) were with Parambassis ranga GSTA-4. Protein rHaGSTA-4 exhibited the highest conjugation activity towards 1-chloro-2,4-dinitrobenzene (CDNB) at pH 7 and 20 ◦C. A disk diffusion assay showed that rHaGSTA-4 significantly protects cells from the stress of exposure to ROS inducers such as CuSO4, CdCl2, and ZnCl2. Furthermore, overexpressed HaGSTA-4 defended cells against oxidative stress caused by H2O2; evidence of selenium-independent peroxidase activity. From qPCR, the tissue-specific expression profile demonstrates that HaGSTA-4 is most highly expressed in the kidney, followed by the intestine and stomach, among fourteen different tissues extracted from healthy seahorses. The mRNA expression profile of HaGSTA-4 upon immune challenge varied depending on the tissue and the time after challenge. Altogether, this study suggests that HaGSTA-4 may be involved in protection against oxidative stress, in immune defense regulation, and xenobiotic metabolism.

목차

ABSTRACT
1. Introduction
2. Materials and methods
2.1. HaGSTA-4 sequence analysis
2.2. Rearing of seahorses and tissue collection
2.3. Immune challenge experiment with bacterial pathogens and immune stimulants
2.4. RNA isolation and cDNA synthesis
2.5. Quantitative real-time PCR (qPCR) for analyzing the spatial and temporal expression of HaGSTA-4
2.6. Preparation of recombinant HaGSTA-4 plasmid constructs
2.7. Recombinant protein (rHaGSTA-4-MBP) expression and purification
2.8. The HaGSTA-4 - pcDNA™3.1(+) vector construction, cell culture, and transfection
2.9. Functional characterization of rHaGSTA-4
2.10. Statistical analysis
3. Results
3.1. HaGSTA-4 sequence analysis
3.2. Tissue-expression pattern of HaGSTA-4
3.3. Temporal expression of HaGSTA-4 after immune challenge
3.4. Induction and purification of rHaGSTA-4
3.5. Specific activities and kinetics of recombinant HaGSTA-4
3.6. Effect of environmental variables (pH and temperature) on rHaGSTA-4 activity
3.7. The antioxidant activity of rHaGSTA-4
3.8. Cell protective role of HaGSTA-4 against H2O2 toxicity
4. Discussion
5. Conclusion
References

키워드

Hippocampus abdominalis Glutathione-S-transferase Alpha-4 Xenobiotic metabolism Immune response

저자

  • Kishanthini Nadarajapillai [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province 63243, Republic of Korea ]
  • D.S. Liyanage [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province 63243, Republic of Korea ]
  • Sarithaa Sellaththurai [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province 63243, Republic of Korea; Marine Science Institute, Jeju National University, Jeju Self-Governing Province 63333, Republic of Korea ]
  • Taehyug Jeong [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province 63243, Republic of Korea; Marine Science Institute, Jeju National University, Jeju Self-Governing Province 63333, Republic of Korea ]
  • Sukkyoung Lee [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province 63243, Republic of Korea; Marine Science Institute, Jeju National University, Jeju Self-Governing Province 63333, Republic of Korea ] Corresponding Author
  • Jehee Lee [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province 63243, Republic of Korea; Marine Science Institute, Jeju National University, Jeju Self-Governing Province 63333, Republic of Korea ] Corresponding Author

참고문헌

자료제공 : 네이버학술정보

간행물 정보

발행기관

  • 발행기관명
    제주대학교 해양과학연구소 [Marine Science Institute Jeju National University]
  • 설립연도
    1968
  • 분야
    농수해양>수산학
  • 소개
    해양의 실체와 그 자원의 합리적 이용과 관리방법을 규명하기 위하여 해양물리, 화학, 생물, 지질, 공학등 여러 관련분야에 관한 다양한 연구를 수행하며, 환경의 보전과 보호를 위해서 수질, 대기, 상·하수도, 폐기물, 사회적 환경 등의 연구를 통하여 쾌적하고 청정한 환경을 보전하기 위한 다양한 연구를 수행하고, 또한 해양 및 환경분야의 우수한 인력양성, 국내외 학술교류의 증진, 학·산 협동체제의 확립 등을 통하여 해양과 환경에 관한 기초 및 응용적 학술연구를 종합적으로 수행함을 목적으로 한다.

간행물

  • 간행물명
    해양과학연구소 연구논문집 [Bulletin of the Marine Science Institute]
  • 간기
    폐간
  • pISSN
    1225-5734
  • 수록기간
    1977~2021
  • 십진분류
    KDC 454 DDC 551

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