Therapeutic Cationic Antimicrobial Peptide (CAP) Derived from Fish Aspartic Proteinase Cathepsin D and its Antimicrobial Mechanism
- 172 Downloads
Aquatic organisms are rich in antimicrobial peptides which play key role against pathogens during infections. Cathepsin is one of immune proteases which have been proven with multiple functions including antimicrobial activity, but their role as antimicrobial peptides has not been elucidated so far in aquatic organisms. This study reports the identification and characterization of antimicrobial peptides from fish cathepsin D. Channa striatus (Cs) cathepsin D (Cath D) was identified from its established cDNA library. Multiple sequence alignment was performed to analyze the homology of CsCathD with other cathepsin. Based on the amino acid propensity scale, two putative antimicrobial regions were identified, synthesized and analyzed for their antimicrobial potency. Gene expression of CsCath D and its mRNA pattern upon pathogenic infection was also observed using real time PCR. All the bioinformatics analysis indicated the gene specific characteristic features of CsCath D. CsCathD mRNA expression was highly expressed at 24 h for bacteria (Aeromonas hydrophila) and 48 h for fungus (Aphanomyces invadans). The CsCath D derived CAPs namely, PL12 and NM12 showed their commendable inhibition towards Bacillus mycoides of the tested bacteria. The cell membrane disruption was observed with PL12 against B. mycoides in flow cytometer. With all proceedings, it is possible to conclude that CsCathD might be a potent immuno modulator and the reported CAPs could be developed as therapeutic agents to treat bacterial pathogenic infections.
KeywordCationic anti-microbial peptide Fish Gene expression Cell membrane disruption Flow cytometry
- Cath D
Open reading frame
Cationic antimicrobial peptides
Epizootic ulcerative syndrome
Minimum inhibitory concentration
Red blood cells
- qRT PCR
Quantitative real time polymerase chain reaction
American type culture collection
Microbial type culture collection
This research is supported by Department of Biotechnology (DBT), Ministry of Science and Technology, Government of India, New Delhi through DBT’s Prestigious Ramalingaswami Re-entry Fellowship (BT/RLF/Re-entry/27/2011). The corresponding author is grateful to the Deanship of Scientific Research, King Saud University for partial funding through Vice Deanship of Scientific Research Chairs. Also, the corresponding author would like to acknowledge Universiti Putra Malaysia, Malaysia for providing him visiting professor award (UPM/PEND/500-3/4/10) to complete this study under the HICoE Program, Ministry of Higher Education Malaysia.
This study was funded by Department of Biotechnology (BT/RLF/Re-entry/27/2011), Ministry of Science and Technology, Government of India, New Delhi; Deanship of Scientific Research, King Saud University and Universiti Putra Malaysia (UPM/PEND/500-3/4/10).
Compliance with Ethical Standards
Conflict of interest
Akila Sathyamoorthi, Venkatesh Kumaresan, Rajesh Palanisamy, Mukesh Pasupuleti, Mariadhas Valan Arasu, Naif Abdullah Al-Dhabi, Kasi Marimuthu, S. M. Nurul Amin, Aziz Arshad, Fatimah Md. Yusoff and Jesu Arockiaraj declare that they have no conflict of interest.
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional [SRM University (361/IEC/2012) and CSIR-CDRI, Lucknow (CDRI/IEC/2014/A1)] and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.
Informed consent was obtained from all individual participants included in the study.
- Arockiaraj J, Haniffa MA, Seetharaman S, Perumalsamy PRR (2004) Utilization of lipid as dietary energy source for fingerlings of Channa striatus. Malays J Sci 23(2):1–5Google Scholar
- Baechle D, Flad T, Cansier A, Steffen H, Schittek B, Tolson J, Herrmann T, Dihazi H, Beck A, Mueller GA, Mueller M, Stevanovic S, Garbe C, Mueller CA, Kalbacher H (2006) Cathepsin D is present in human eccrine sweat and involved in the postsecretory processing of the antimicrobial peptide DCD-1L. J Biol Chem 281(9):5406–5415CrossRefGoogle Scholar
- Ellis AE (1982) Differences between the immune mechanisms of fish and higher vertebrates. In: Robert R (ed) Microbial disease of fish. Academic Press, London, pp 1.30Google Scholar
- Jayaram KC (1981) Freshwater fishes of India, Pakistan, Bangladesh, Burma and Srilanka. A handbook, 1st edn. Zoological Survey of India, CalcuttaGoogle Scholar
- Khurshid MA (1999) Staphylococcus aureus with reduced susceptibility to vancomycin. Morbid Mort Week Rep 48:1165–1167Google Scholar
- Koplan JP (1998) Preventing emerging infectious diseases. A strategy for the 21st century. US Department of Health and Human Services, Centers for Disease Control and Prevention, AtlantaGoogle Scholar
- Lilley JH, Callinan RB, Chinabut S, Kanchanakhan S, MacRae IH, Phillips MJ (1998) Epizootic ulcerative syndrome (EUS) technical handbook. The Aquatic Animal Health Research Institute, Bangkok, p 88Google Scholar
- Pan CY, Chen JY, Cheng YS, Chen CY, Ni IH, Sheen JF, Pan YL, Kuo CM (2007) Gene expression and localization of the epinecidin-1 antimicrobial peptide in the grouper (Epinephelus coioides), and its role in protecting fish against pathogenic infection. DNA Cell Biol 26(6):403–413CrossRefGoogle Scholar