Identification of the first cathelicidin gene from skin of Chinese giant salamanders Andrias davidianus with its potent antimicrobial activity

• DOI: 10.1016/j.dci.2017.08.002
• Publication Date: 2017-08-09T17:45:15Z
• Authors: Hui Yang; Baoyue Lu; Dandan Zhou; Lin Zhao; Weijia Song; Lixin Wang
• Keywords: Lipopolysaccharides; 0301 basic medicine; Urodela; Microbial Sensitivity Tests; Amphibian Proteins; Immunity, Innate; Recombinant Proteins; Cell Line; 3. Good health; 03 medical and health sciences; Bacteriolysis; Anti-Infective Agents; Cathelicidins; Animals; Cloning, Molecular; Transcriptome; Protein Binding; Skin

Cathelicidins, as effector molecules, play important roles against infections and represent a crucial component of the innate immune system in vertebrates. They are widely studied in mammals, but little is known in amphibians. In the present study, we report the identification and characterization of a novel cathelicidin from Chinese giant salamander Andrias davidianus, which is the first study in Caudata amphibian. The cDNA sequence encodes a predicted 148-amino-acid polypeptide, which composed of a 20-residue signal peptide, a 94-residue conserved cathelin domain and a 34-residue mature peptide. From the multiple sequence alignments and phylogenetic analysis, AdCath shared conserved structure with other orthologs and clustered with other amphibian peptides. The tissue expression profiles revealed AdCath was highly expressed in skin. To study the function of AdCath gene, the AdCath precursor protein and mature peptide were recombinantly expressed and chemical synthesized respectively. The rAdCath protein could bind to LPS in a dose-dependent manner. When the concentrations of rAdCath protein and mature peptide were up to 22 μg/mL, they showed significantly cytotoxicity to human 293T cell lines. The rAdCath protein and synthetic peptide could exhibit antibacterial activities detected by the minimum inhibitory concentrations assay. From the SEM assay, the synthetic mature peptide could destroy the membrane of bacteria and cause loss of membrane integrity. Collectively, these findings characterized the first cathelicidin from A. davidianus, and highlighted its potential antimicrobial activities, indicating its important roles in the skin immune response against different bacteria.