Molecular characterization and phylogenetic analysis of a penaeidin-like antimicrobial peptide, Fi-penaeidin from Fenneropenaeus indicus
Introduction
Cationic antimicrobial peptides (AMPs) play important roles as effectors of the first line of defense in innate immunity of every animal, due to their broad-spectrum antimicrobial activity and rapid production. Many AMPs show a remarkable specificity for prokaryotes with low toxicity for eukaryotic cells; a phenomenon which has favored their investigation and exploitation as potential new antibiotics. In penaeid shrimps, three main families of AMPs have been currently described: anti-lipopolysaccharide factors (ALFs), crustins and penaeidins. ALFs possess a broader antimicrobial spectrum (Somboonwiwat et al., 2005), whereas, crustins are reported to have a more-restricted activity spectrum, affecting mainly marine Gram-positive bacteria (Relf et al., 1999). Penaeidins, first characterized from Litopenaeus vannamei, act mainly against Gram-positive bacteria, filamentous fungi (Destoumieux et al., 1997), viruses and protozoans (Bachere, 2003) and are also found to possess chitin-binding properties (Destoumieux et al., 2000). Recent studies have revealed the presence of penaeidin mRNAs in different penaeid shrimp species viz. L. vannamei (Destoumieux et al., 1997); Litopenaeus setiferus (Gross et al., 2001); Penaeus monodon (Supungul et al., 2002); Penaeus japonicus (Rojtinnakorn et al., 2002); Litopenaeus schmitti (Barracco et al., 2005); Fenneropenaeus chinensis (Kang et al., 2004); Farfantepenaeus paulensis (Barracco et al., 2005) and Litopenaeus stylirostris (Munoz et al., 2004). Gueguen et al. (2006) have developed a penaeidin database, named as ‘PenBase’ to provide comprehensive information about penaeidin properties, diversity and nomenclature.
The Indian white shrimp, Fenneropenaeus indicus is the most dominant shrimp species in the shallow water shrimp fishery along the west and east coast of India and an important shrimp species which is cultivated in Pokkali paddy fields (paddy-cum-prawn farming) practiced in the wetlands of Kerala. Though several research works have been carried out for unraveling the biochemical aspects of the defense mechanisms in F. indicus, few works have been carried out on AMPs in this species. So far no penaeidin sequences have been reported from F. indicus. Discovery of new penaeidin isoforms in F. indicus will certainly help us to unravel the host defense mechanism of this shrimp species. In the current study, a penaeidin cDNA has been characterized from the Indian white shrimp, F. indicus. This is the first report of this peptide molecule from F. indicus.
Section snippets
Experimental animals and haemolymph collection
Healthy adult F. indicus (8–10 g body weight) were collected from a local shrimp farm in Vypeen, Kochi. They were transferred to aquarium tanks and maintained under laboratory conditions. Haemolymph was collected from the rostral sinus using specially designed capillary tubes (RNase-free) and rinsed using pre-cooled anticoagulant solution (RNase free 10% sodium citrate, pH 7.0). Haemolymph was suspended in TRI reagent (Sigma) for total RNA isolation.
Total RNA isolation and reverse transcription
Total RNA was extracted from the haemocytes
Results and discussion
A 263 bp fragment of PCR amplified product was obtained from the mRNA of F. indicus haemocytes by RT-PCR (Fig. 1A). The penaeidin cDNA was found to consist of 186 bp nucleotides, encoding 61 amino acids and an ORF of 42 amino acids. The ORF encoded 42 amino acid residues with a predicted molecular weight (MW) of 4.478 kDa and theoretical isoelectric point (pI) of 5.3 as calculated by the PROTPARAM software. The analysis with the SignalP software revealed the presence of a signal peptide with 19
Conclusion
The discovery of AMPs in crustaceans provides new clues for fundamental understanding of crustacean immunity. The studies on the shrimp penaeidins have largely contributed to this knowledge as they are the first AMPs fully characterized in crustaceans. This is the first report of penaeidin AMP from Indian white shrimp, F. indicus. The wide distribution of penaeidins in penaeid shrimps indicates the importance of these AMPs in the innate immunity and a detailed research is required to reveal the
Acknowledgment
The authors are grateful to the Ministry of Earth Sciences (MoES), Govt. of India for the research grant (MoES/10-MLR/2/2007) with which the work was carried out. The first author gratefully acknowledges KSCSTE (Kerala State Council for Science, Technology and Environment) for the award of fellowship.
References (13)
Anti-infectious immune effectors in marine invertebrates, potential tools for disease control in larviculture
Aquaculture.
(2003)- et al.
Molecular characterization of penaeidins from two Atlantic brazilian shrimp species
Farfantepenaeus paulensis and Litopenaeus schmitti. FEMS Microbiology Letters.
(2005) - et al.
Penaeidins, a new family of antimicrobial peptides isolated from the shrimp Penaeus vannamei (Decapoda)
J. Biol. Chem.
(1997) - et al.
Immune gene discovery by expressed sequence tag analysis of hemocytes and hepatopancreas in the Pacific white shrimp, Litopenaeus vannamei, and Atlantic white shrimp, Litopenaeus setiferus
Dev. Comp. Immunol.
(2001) - et al.
PenBase, the shrimp antimicrobial peptide penaeidin database, Sequence-based classification and recommended nomenclature
Dev. Comp. Immunol.
(2006) - et al.
Molecular cloning and expression analysis of Ch-penaeidin, an antimicrobial peptide from Chinese shrimp, Fenneropenaeus chinensis
Fish Shellfish Immunol.
(2004)
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