Molecular dynamics simulation studies and in vitro site directed mutagenesis of avian beta-defensin Apl_AvBD2

BMC Bioinformatics

Research

Krishnankutty Chandrika Sivakumar¹  Easwaran Sreekumar²  Soja Saghar Soman² 

[1] Bioinformatics facility, Rajiv Gandhi Centre for Biotechnology (RGCB), Thycaud PO, 695014, Thiruvananthapuram, Kerala, India;Molecular Virology Laboratory, Department of Molecular Microbiology, Rajiv Gandhi Centre for Biotechnology (RGCB), Thycaud PO, 695014, Thiruvananthapuram, Kerala, India;

关键词: Molecular Dynamic Simulation;    Antimicrobial Peptide;    Root Mean Square Deviation;    Homology Modelling;    Arginine Residue;   

DOI  :  10.1186/1471-2105-11-S1-S7

来源: Springer

【 摘 要 】

BackgroundDefensins comprise a group of antimicrobial peptides, widely recognized as important elements of the innate immune system in both animals and plants. Cationicity, rather than the secondary structure, is believed to be the major factor defining the antimicrobial activity of defensins. To test this hypothesis and to improve the activity of the newly identified avian β-defensin Apl_AvBD2 by enhancing the cationicity, we performed in silico site directed mutagenesis, keeping the predicted secondary structure intact. Molecular dynamics (MD) simulation studies were done to predict the activity. Mutant proteins were made by in vitro site directed mutagenesis and recombinant protein expression, and tested for antimicrobial activity to confirm the results obtained in MD simulation analysis.ResultsMD simulation revealed subtle, but critical, structural variations between the wild type Apl_AvBD2 and the more cationic in silico mutants, which were not detected in the initial structural prediction by homology modelling. The C-terminal cationic 'claw' region, important in antimicrobial activity, which was intact in the wild type, showed changes in shape and orientation in all the mutant peptides. Mutant peptides also showed increased solvent accessible surface area and more number of hydrogen bonds with the surrounding water molecules. In functional studies, the Escherichia coli expressed, purified recombinant mutant proteins showed total loss of antimicrobial activity compared to the wild type protein.ConclusionThe study revealed that cationicity alone is not the determining factor in the microbicidal activity of antimicrobial peptides. Factors affecting the molecular dynamics such as hydrophobicity, electrostatic interactions and the potential for oligomerization may also play fundamental roles. It points to the usefulness of MD simulation studies in successful engineering of antimicrobial peptides for improved activity and other desirable functions.

【 授权许可】

Unknown   
© Soman et al; licensee BioMed Central Ltd. 2010. This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.