Abstract
Snakins are antimicrobial peptides (AMPs) found, so far, exclusively in plants, and known to be important in the defense against a wide range of pathogens. Like other plant AMPs, they contain several positively charged amino acids, and an even number of cysteine residues forming disulfide bridges which are considered important for their usual function. Despite its importance, studies on snakin tertiary structure and mode of action are still scarce. In this study, a new snakin-like gene was isolated from the native plant Peltophorum dubium, and its expression was verified in seedlings and adult leaves. The deduced peptide (PdSN1) shows 84% sequence identity with potato snakin-1 mature peptide, with the 12 cysteines characteristic from this peptide family at the GASA domain. The mature PdSN1 coding sequence was successfully expressed in Escherichia coli. The purified recombinant peptide inhibits the growth of important plant and human pathogens, like the economically relevant potato pathogen Streptomyces scabies and the opportunistic fungi Candida albicans and Aspergillus niger. Finally, homology and ab initio modeling techniques coupled to extensive molecular dynamics simulations were used to gain insight on the 3D structure of PdSN1, which exhibited a helix–turn–helix motif conserved in both native and recombinant peptides. We found this motif to be strongly coded in the sequence of PdSN1, as it is stable under different patterns of disulfide bonds connectivity, and even when the 12 cysteines are considered in their reduced form, explaining the previous experimental evidences.
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Acknowledgements
The authors thank CSIC (Comisión Sectorial de Investigación Científica, Universidad de la República, Uruguay), PEDECIBA (Programa de Desarrollo de las Ciencias Básicas, Uruguay), ANII (Agencia Nacional de Investigación e Innovación, Uruguay), CNPq (National Council for Scientific and Technological Development, Brazil), and CAPES (Coordination for the Improvement of Higher Education Personnel, Brazil) for financial support and fellowships. We also thank Dr. Rosario Durán and Madelón Portela for MALDI-TOFF analysis. SRD was supported by a doctoral fellowship from CSIC. IRB Barcelona is the recipient of a Severo Ochoa Award of Excellence from MINECO (Ministerio de Economía, Industria y Competitividad. Government of Spain). PDD and GC are PEDECIBA and SNI (Sistema Nacional de Investigadores, Uruguay) researchers.
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Rodríguez-Decuadro, S., Barraco-Vega, M., Dans, P.D. et al. Antimicrobial and structural insights of a new snakin-like peptide isolated from Peltophorum dubium (Fabaceae). Amino Acids 50, 1245–1259 (2018). https://doi.org/10.1007/s00726-018-2598-3
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DOI: https://doi.org/10.1007/s00726-018-2598-3