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Identification of a novel cathelicidin antimicrobial peptide from ducks and determination of its functional activity and antibacterial mechanism.

Gao W, Xing L, Qu P, Tan T, Yang N, Li D, Chen H, Feng X - Sci Rep (2015)

Bottom Line: The cDNA sequence of dCATH encodes a predicted 146-amino-acid polypeptide composed of a 17-residue signal peptide, a 109-residue conserved cathelin domain and a 20-residue mature peptide.Phylogenetic analysis demonstrated that dCATH is highly divergent from other avian peptides.The effects on bacterial outer and inner membranes, as examined by scanning electron microscope and transmission electron microscopy, indicate that dCATH kills microbial cells by increasing permeability, causing a loss of membrane integrity.

View Article: PubMed Central - PubMed

Affiliation: College of Animal Science and Technology, Northeast Agricultural University, Harbin 150030, P.R. China.

ABSTRACT
The family of antimicrobial peptide, cathelicidins, which plays important roles against infections in animals, has been identified from many species. Here, we identified a novel avian cathelicidin ortholog from ducks and named dCATH. The cDNA sequence of dCATH encodes a predicted 146-amino-acid polypeptide composed of a 17-residue signal peptide, a 109-residue conserved cathelin domain and a 20-residue mature peptide. Phylogenetic analysis demonstrated that dCATH is highly divergent from other avian peptides. The α-helical structure of the peptide exerted strong antimicrobial activity against a broad range of bacteria in vitro, with most minimum inhibitory concentrations in the range of 2 to 4 μM. Moreover, dCATH also showed cytotoxicity, lysing 50% of mammalian erythrocytes in the presence or absence of 10% fetal calf serum at concentrations of 32 μM or 20 μM and killing 50% HaCaT cells at a concentration of 10 μM. The effects on bacterial outer and inner membranes, as examined by scanning electron microscope and transmission electron microscopy, indicate that dCATH kills microbial cells by increasing permeability, causing a loss of membrane integrity.

No MeSH data available.


Related in: MedlinePlus

Nucleotide and amino acid sequences for the peptide.Start and stop codons are bold. Signal peptide sequence predicted by SignalP Version 3.0 software (http://www.cbs.dtu.dk/services/SignalP/) is underlined, the four cysteines are bold and underlined. The predicted mature peptide is displayed in bold and italics.
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f1: Nucleotide and amino acid sequences for the peptide.Start and stop codons are bold. Signal peptide sequence predicted by SignalP Version 3.0 software (http://www.cbs.dtu.dk/services/SignalP/) is underlined, the four cysteines are bold and underlined. The predicted mature peptide is displayed in bold and italics.

Mentions: As shown in Fig. 1, one positive clone containing the dCATH cathelicidin cDNA was identified and isolated. The 441-bp cDNA and the deduced 146-amino-acid sequence from the cDNA sequence are shown in Fig. 1, which has been deposited in GenBank (GenBank accession number: KT230679). The N-terminal region contained a classical 17-amino-acid N-terminal signal peptide sequence, as predicted by SignalP software (Version 3.0, http://www.cbs.dtu.dk/services/SignalP/). The remaining propeptide contained a characteristic cathelin-like domain with two conserved cysteine bridges and a C-terminal antimicrobial domain. Given that valine is the optimal protease cleavage site for the maturation of most cathelicidins1617, valine at position 126 (Val126–Lys127) is assumed to be the elastase cleavage site, which releases the mature dCATH: KRFWQLVPLAIKIYRAWKRR.


Identification of a novel cathelicidin antimicrobial peptide from ducks and determination of its functional activity and antibacterial mechanism.

Gao W, Xing L, Qu P, Tan T, Yang N, Li D, Chen H, Feng X - Sci Rep (2015)

Nucleotide and amino acid sequences for the peptide.Start and stop codons are bold. Signal peptide sequence predicted by SignalP Version 3.0 software (http://www.cbs.dtu.dk/services/SignalP/) is underlined, the four cysteines are bold and underlined. The predicted mature peptide is displayed in bold and italics.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4660463&req=5

f1: Nucleotide and amino acid sequences for the peptide.Start and stop codons are bold. Signal peptide sequence predicted by SignalP Version 3.0 software (http://www.cbs.dtu.dk/services/SignalP/) is underlined, the four cysteines are bold and underlined. The predicted mature peptide is displayed in bold and italics.
Mentions: As shown in Fig. 1, one positive clone containing the dCATH cathelicidin cDNA was identified and isolated. The 441-bp cDNA and the deduced 146-amino-acid sequence from the cDNA sequence are shown in Fig. 1, which has been deposited in GenBank (GenBank accession number: KT230679). The N-terminal region contained a classical 17-amino-acid N-terminal signal peptide sequence, as predicted by SignalP software (Version 3.0, http://www.cbs.dtu.dk/services/SignalP/). The remaining propeptide contained a characteristic cathelin-like domain with two conserved cysteine bridges and a C-terminal antimicrobial domain. Given that valine is the optimal protease cleavage site for the maturation of most cathelicidins1617, valine at position 126 (Val126–Lys127) is assumed to be the elastase cleavage site, which releases the mature dCATH: KRFWQLVPLAIKIYRAWKRR.

Bottom Line: The cDNA sequence of dCATH encodes a predicted 146-amino-acid polypeptide composed of a 17-residue signal peptide, a 109-residue conserved cathelin domain and a 20-residue mature peptide.Phylogenetic analysis demonstrated that dCATH is highly divergent from other avian peptides.The effects on bacterial outer and inner membranes, as examined by scanning electron microscope and transmission electron microscopy, indicate that dCATH kills microbial cells by increasing permeability, causing a loss of membrane integrity.

View Article: PubMed Central - PubMed

Affiliation: College of Animal Science and Technology, Northeast Agricultural University, Harbin 150030, P.R. China.

ABSTRACT
The family of antimicrobial peptide, cathelicidins, which plays important roles against infections in animals, has been identified from many species. Here, we identified a novel avian cathelicidin ortholog from ducks and named dCATH. The cDNA sequence of dCATH encodes a predicted 146-amino-acid polypeptide composed of a 17-residue signal peptide, a 109-residue conserved cathelin domain and a 20-residue mature peptide. Phylogenetic analysis demonstrated that dCATH is highly divergent from other avian peptides. The α-helical structure of the peptide exerted strong antimicrobial activity against a broad range of bacteria in vitro, with most minimum inhibitory concentrations in the range of 2 to 4 μM. Moreover, dCATH also showed cytotoxicity, lysing 50% of mammalian erythrocytes in the presence or absence of 10% fetal calf serum at concentrations of 32 μM or 20 μM and killing 50% HaCaT cells at a concentration of 10 μM. The effects on bacterial outer and inner membranes, as examined by scanning electron microscope and transmission electron microscopy, indicate that dCATH kills microbial cells by increasing permeability, causing a loss of membrane integrity.

No MeSH data available.


Related in: MedlinePlus