Molecular Characterisation of a Novel Isoform of Hepatic Antimicrobial Peptide, Hepcidin (Le-Hepc), from Leiognathus equulus and Analysis of Its Functional Properties In Silico
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Hepcidin represents a family of cysteine-rich antimicrobial peptides that are mainly expressed in the liver of living organisms. In this study, we have identified and characterised a novel isoform of hepcidin from the common pony fish, Leiognathus equulus (Le-Hepc). A 261-bp fragment cDNA coding for 86 amino acids was obtained. Homologous analysis showed that Le-Hepc belongs to the hepcidin super family and shares sequence identity with other known fish pre-propeptide hepcidin sequences. The ORF encodes for a 24-amino acid (aa) signal peptide coupled to a 36-aa prodomain followed by a 26-aa mature peptide. The mature peptide region has a calculated molecular weight of 2.73 kDa, a net positive charge of +2 and a theoretical pI of 8.23. Phylogenetic analysis of Le-Hepc showed a strong relationship with other fish hepcidin sequences and clustered into HAMP2 group hepcidins. Secondary structural analysis indicated that Le-Hepc mature peptide contains two antiparallel β-sheets strengthened by four disulphide bonds formed by eight conserved cysteine residues. The physicochemical properties of the peptide and its structural parameters are in agreement with characteristic features of an antimicrobial peptide. This is the first report of an antimicrobial peptide from the common pony fish, L. equulus.
KeywordsAntimicrobial peptide Hepcidin Pony fish Leiognathus equulus HAMP2
The authors are grateful to the Director, Centre for Marine Living Resources and Ecology (CMLRE) and Ministry of Earth Sciences (MoES), Govt. of India, for the research grant (MoES/10-MLR/01/2012) and scientific support for the work. The authors also thank the Coordinator, National Centre for Aquatic and Animal Health (NCAAH), for scientific support. The first author gratefully acknowledges UGC-BSR (University Grant Commission-Basic Scientific Research) for the award of a fellowship.
Compliance with Ethical Standards
The study was funded by the Ministry of Earth Sciences (MoES), Govt. of India (MoES/10-MLR/01/2012).
Conflict of Interest
Aishwarya Nair, Sruthy K. S., Chaithanya E.R., Sajeevan T.P., I. S. Bright Singh and Rosamma Philip declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- 18.Liang T, Ji W, Zhang GR, Wei KJ, Feng K, Wang WM, Zou GW (2013) Molecular cloning and expression analysis of liver-expressed antimicrobial peptide 1 (LEAP-1) and LEAP-2 genes in the blunt snout bream(Megalobrama amblycephala). Fish Shellfish Immunol 35:553–563. doi: 10.1016/j.fsi.2013.05.021 CrossRefGoogle Scholar
- 21.Chi JR, Liao LS, Wang RG, Jhu CS, Wu JL, Hu SY (2014) Molecular cloning and functional characterization of the hepcidin gene from the convict cichlid (Amatitlania nigrofasciata) and its expression pattern in response to lipopolysaccharide challenge. Fish Physiol Biochem 41:449–461. doi: 10.1007/s10695-014-9996-6 CrossRefGoogle Scholar
- 28.Lauth X, Babon JJ, Stannard JA, Singh S, Nizet V, Carlberg JM, Ostland VE, Pennington MW, Norton RS, Westerman ME (2005) Bass hepcidin synthesis, solution structure, antimicrobial activities and synergism, and in vivo hepatic response to bacterial infections. J Biol Chem 280(10):9272–9282. doi: 10.1074/jbc.M411154200 CrossRefGoogle Scholar
- 32.Chaithanya ER, Philip R, Sathyan N, Anil Kumar PR, Antony SP, Sanjeevan VN, Bright Singh IS (2013) A novel isoform of the hepatic antimicrobial peptide, hepcidin (Hepc-CB1), from a deep-sea fish, the spinyjaw greeneye Chlorophthalmus bicornis (Norman, 1939): molecular characterisation and phylogeny. Probiotics Antimicrob Proteins 5(1):1–7. doi: 10.1007/s12602-012-9120-0 CrossRefGoogle Scholar
- 46.Chekmenev EY, Jones SM, Nikolayeva YN, Vollmar BS, Wagner TJ, Gor'kov PL, Brey WW, Manion MN, Daugherty KC, Cotten M (2006) High-field NMR studies of molecular recognition and structure-function relationships in antimicrobial piscidins at the water-lipid bilayer interface. J Am Chem Soc 128:5308–5309. doi: 10.1021/ja058385e CrossRefGoogle Scholar
- 47.Lynch M, O’Hely M, Walsh B, Force A (2001) The probability of preservation of a newly arisen gene duplicate. Genetics 159:1789–1804Google Scholar