Earticle

초록

영어

Malectin is a carbohydrate-binding lectin protein found in the endoplasmic reticulum (ER). It selectivity binds to Glc2-N-glycan and is involved in a glycoprotein quality control mechanism. Even though malectin may play a role in immunity, its role in innate immunity is not fully known. In the present study, we identified and characterized the malectin gene from Hippocampus abdominalis (HaMLEC). We analyzed sequence features, spatial expression levels, temporal expression profiles upon immune responses, bacterial and carbohydrate binding abilities and anti-viral properties to investigate the potential role of HaMLEC in innate immunity. The molecular weight and isoelectric point (pI) were estimated to be 31.99 kDa and 5.17, respectively. The N-terminal signal peptide, malectin superfamily domain and C-terminal transmembrane region were identified from the amino acid sequence of HaMLEC. The close evolutionary relationship of HaMLEC with other teleosts was identified by phylogenetic analysis. According to quantitative PCR (qPCR) results, HaMLEC expression was observed in all the examined tissues and high expression was observed in the ovary and brain, compared to other tested tissues. Temporal expression of HaMLEC in liver and blood tissues were significant modulated upon exposure to immunogens Edwardasiella tarda, Streptococcus iniae, polyinosinic:polycytidylic and lipopolysaccharide. The presence of carbohydrate binding modules (CBMs) of bacterial glycosyl hydrolases were functionally confirmed by a bacterial binding assay. Anti-viral activity significantly reduced viral hemorrhagic septicemia virus (VHSV) replication in cells overexpressing HaMLEC. The observed results suggested that HaMLEC may have a significant role in innate immunity in Hippocampus abdominalis.

목차

ABSTRACT
1. Introduction
2. Materials and methods
2.1. Identification of seahorse malectin cDNA sequences
2.2. Molecular profiling of HaMLEC using bioinformatic tools
2.3. Acclimatization of experimental seahorse and tissue sampling
2.4. Immune challenge experiment
2.5. Total RNA extraction and cDNA synthesis
2.6. Transcriptional analysis of HaMLEC by qRT-PCR
2.7. Construction of eukaryotic and prokaryotic expression vectors
2.8. Prokaryotic overexpression and purification of rHaMLEC protein
2.9. Bacterial agglutination activity of rHaMLEC
2.10. Microbial binding activity assay
2.11. Cell culture and transfection
2.12. Antiviral activity assay
2.13. Statistical analysis
3. Results
3.1. Comprehensive molecular characterization of HaMLEC
3.2. Spatial mRNA expression of HaMLEC
3.3. Analysis of HaMLEC expression levels after immune challenge
3.4. Overexpression and purification of rHaMLEC
3.5. Functional protein studies
4. Discussion
CRediT authorship contribution statement
Acknowledgments
References

키워드

저자

• 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 ]
• K.A.S.N. Shanaka [ 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 ]
• D.S. Liyanage [ 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 ]
• Hyerim Yang [ 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 ]
• Thanthrige Thiunuwan Priyathilaka [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province, 63243, Republic of Korea ]
• 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

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  해양과학연구소 연구논문집 [Bulletin of the Marine Science Institute]

• 간기

  폐간

• pISSN

  1225-5734

• 수록기간

  1977~2021

• 십진분류

  KDC 454 DDC 551