Recombinant expression and solution structure of antimicrobial peptide aurelin from jellyfish Aurelia aurita

• Published in: Biochemical and biophysical research communications 2012-12, Vol.429 (1-2), p.63-69
• Main Authors: Shenkarev, Zakhar O., Panteleev, Pavel V., Balandin, Sergey V., Gizatullina, Albina K., Altukhov, Dmitry A., Finkina, Ekaterina I., Kokryakov, Vladimir N., Arseniev, Alexander S., Ovchinnikova, Tatiana V.
• Format: Article
• Language: English
• Subjects: 60 APPLIED LIFE SCIENCES; Amino Acid Sequence; Animals; Antimicrobial Cationic Peptides - biosynthesis; Antimicrobial Cationic Peptides - chemistry; Antimicrobial Cationic Peptides - pharmacology; Antimicrobial peptide; Aurelia aurita; Aurelin; Bacteria - drug effects; BERNSTEIN MODE; DETERGENTS; DISULFIDES; ESCHERICHIA COLI; LIPIDS; Marine; Micelles; Molecular Sequence Data; NMR; NUCLEAR MAGNETIC RESONANCE; PARAMAGNETISM; PEPTIDES; Phosphatidylcholines - chemistry; Phosphatidylglycerols - chemistry; POTASSIUM IONS; Protein Structure, Secondary; Recombinant expression; Recombinant Proteins - biosynthesis; Recombinant Proteins - chemistry; Recombinant Proteins - pharmacology; Scyphozoa; SEAS; Solutions; Spatial structure; TOXINS; Water - chemistry; ZWITTERIONIC COMPOUNDS
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2012.10.092

[Display omitted] ► Aurelin was overexpressed in Escherichia coli, and its spatial structure was studied by NMR. ► Aurelin compact structure encloses helical regions cross-linked by three disulfide bonds. ► Aurelin shows structural homology to the BgK and ShK toxins of sea anemones. ► Aurelin binds to the anionic lipid vesicles, but does not interact with zwitterionic ones. ► Aurelin binds to DPC micelle surface with moderate affinity via two helical regions. Aurelin is a 40-residue cationic antimicrobial peptide isolated from the mezoglea of a scyphoid jellyfish Aurelia aurita. Aurelin and its 15N-labeled analogue were overexpressed in Escherichia coli and purified. Antimicrobial activity of the recombinant peptide was examined, and its spatial structure was studied by NMR spectroscopy. Aurelin represents a compact globule, enclosing one 310-helix and two α-helical regions cross-linked by three disulfide bonds. The peptide binds to anionic lipid (POPC/DOPG, 3:1) vesicles even at physiological salt concentration, it does not interact with zwitterionic (POPC) vesicles and interacts with the DPC micelle surface with moderate affinity via two α-helical regions. Although aurelin shows structural homology to the BgK and ShK toxins of sea anemones, its surface does not possess the “functional dyad” required for the high-affinity interaction with the K+-channels. The obtained data permit to correlate the modest antibacterial properties and membrane activity of aurelin.