Piscidin-like peptide

Piscidins, important components of the innate (nonspecific) immunity system in fish, have potent, broad-spectrum antimicrobial and antiparasitic activities. In this study, we reported a novel antimicrobial cationic peptide from Pseudosciaena crocea. Although this peptide exhibited a genomic (3 exons and 2 introns) and propeptide (signal peptide, mature peptide and prodomain) organization, conserved signal peptide (22 amino acids) and consensus motif I-X5-H-X4-I-H identical to the reported fish piscidins, Pc-pis showed a relatively low overall conservation with other known piscidins, which was obviously embodied in the amino acid composition of the peptide. Pc-pis is strikingly rich in glycine residues (27.3%), which disrupted the amphipathic structure of the peptide. Relative quantitative real-time PCR revealed that Pc-pis is a typically gill-expressed peptide. The sequence analysis, structural features and tissue distribution suggested that Pc-pis was genetically related to the piscidins family and might be a novel piscidin-like antimicrobial peptide. Quantitative PCR analysis revealed that the expression of Pc-pis in the spleen, head-kidney, liver, intestine, skin and gill could be regulated during Cryptocaryon irritans infection and post C. irritans falling off, implicating a role for Pc-pis in immune defense against C. irritans and secondary bacterial infections. Synthetic Pc-pis exhibited broad-spectrum activity against bacteria, fungi and C. irritans in parasitic stages. These results provided the first evidence of piscidins antiparasitic activity against marine fish ectoparasites C. irritants trophonts and further indicated that Pc-pis might be an important component of the P. crocea innate immune system against C. irritans and secondary bacterial infections. Thus, these data provided new insights into P. crocea innate immunity against external protozoan parasite and microbial infections and facilitate the evaluation of Pc-pis as a therapeutic agent against pathogen invasion.

Chionodracine (Cnd) is a 22-residue peptide of the piscidin family expressed in the gills of the Chionodraco hamatus as protection from bacterial infections. Here, we report the effects of synthetic Cnd on both Psychrobacter sp. TAD1 and Escherichia coli bacteria, as well as membrane models. We found that Cnd perforates the inner and outer membranes of Psychrobacter sp. TAD1, making discrete pores that cause the cellular content to leak out. Membrane disruption studies using intrinsic and extrinsic fluorescence spectroscopy revealed that Cnd behaves similarly to other piscidins, with comparable membrane partition coefficients. Membrane accessibility assays and structural studies using NMR in detergent micelles show that Cnd adopts a canonical topology of antimicrobial helical peptides, with the hydrophobic face toward the lipid environment and the hydrophilic face toward the bulk solvent. The analysis of Cnd free energy of binding to vesicles with different lipid contents indicates a preference for charged phospholipids and a more marked binding to native E. coli extracts. Taken with previous studies on piscidin-like peptides, we conclude that Cnd first adsorbs to the membrane, and then forms pores together with membrane fragmentation. Since Cnd has only marginal hemolytic activity, it constitutes a good template for developing new antimicrobial agents.

Antimicrobial peptides, which are crucial effectors of innate immunity, are a promising substitute for antibiotics. The piscidin family is a group of fish-derived antimicrobial peptides that have potent antimicrobial activity and participate in the innate immune response. Here we describe a novel piscidin-like peptide called cerocin from the black sea bass (Centropristis striata), which is a highly valued marine teleost in both commercial and recreational fisheries worldwide. The full-length cDNA of cerocin consists of 567 base pairs, including 5' and 3' untranslated regions of 61 and 209 base pairs, respectively. The active peptide consists of 20 amino acids that form an amphipathic α-helix structure. Cerocin showed highest identity with the cardinalfish (Ostorhinchus fasciatus) piscidin (52%). Phylogenetic tree demonstrated that the cerocin clustered with dicentracin of Liparis tanakae and Perca flavescens. It showed tissue-specific distribution patterns and was predominantly expressed in the gill. After challenge with Vibrio harveyi, C. striata showed time- and tissue-dependent expression of the cerocin gene. Finally, a cerocin peptide was synthesized, and it exerted broad-spectrum antimicrobial activity against a number of bacterial strains, especially Gram-positive pathogens. Analysis of the killing kinetics revealed that the cerocin peptide had a rapid bactericidal effect on the bacteria. Collectively, these data suggest that the piscidin-like cerocin might play a vital role in the immune response of C. striata, and further studies of this gene may provide insight into the innate immune system of this species.