‘Lollipop’-shaped helical structure of a hybrid antimicrobial peptide of temporin B-lipopolysaccharide binding motif and mapping cationic residues in antibacterial activity

• Published in: Biochimica et biophysica acta Vol. 1860; no. 6; pp. 1362 - 1372
• Main Authors: Mohanram, Harini, Bhattacharjya, Surajit
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.06.2016
• Subjects: amino acids; Anti-Infective Agents - chemistry; Anti-Infective Agents - pharmacology; antibacterial properties; antibiotics; Antimicrobial peptide; antimicrobial peptides; Beta-boomerang motif; Binding Sites; cations; cell membranes; detergents; Gram-negative bacteria; hydrophobicity; lipopolysaccharides; Lipopolysaccharides - chemistry; Lipopolysaccharides - metabolism; micelles; NMR; nuclear magnetic resonance spectroscopy; Permeability; Protein Structure, Secondary; Proteins - chemistry; Proteins - pharmacology; Structure; Temporin; zwitterions; NMR; Temporin; Structure; Beta-boomerang motif; Antimicrobial peptide
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/j.bbagen.2016.03.025

Temporins are attractive templates for the development of antibiotics. However, many temporins are inactive against Gram-negative bacteria. Previously, we demonstrated conjugation of a lipopolysaccharide binding motif peptide to temporins yielded hybrid non-haemolytic AMPs that killed several Gram-negative bacteria. We carried out a systematic Ala replacement of individual cationic and polar amino acid residues of LG21, a hybrid AMP consisted of temporin B (TB) and LPS binding motif. These Ala containing analogs of LG21 were examined for antibacterial activity, cell membrane permeabilization and liposome leakage assays using optical spectroscopic methods. Atomic resolution structure of LG21 was determined in zwitterionic dodecyl phosphocholine (DPC) micelles by NMR spectroscopy. Cationic residues in the LPS binding motif of LG21 were critical for bactericidal and membrane permeabilization. Detergent bound structure of LG21 revealed helical conformation containing extensive sidechain/sidechain packing including cation/π interactions in the LPS binding motif. The helical structure of LG21 resembled a ‘lollipop’ like shape that was sustained by a compacted bulky aromatic/cationic head with a comparatively thinner ‘stick’ at the N-terminal region. The ‘head’ of the structure could be localized into micelle-water interfacial region whereas the ‘stick’ region may be inserted into the hydrophobic core of micelle. The LPS binding motif of LG21 played dominant roles in broad spectrum activity and the 3-D structure provided plausible mechanistic insights for permeabilization of bacterial membrane. Hybrid AMPs containing LPS binding motif could be useful for the structure based development of broad spectrum antibiotics. •Hybrid AMP LG21 (temporin B+LPS motif) shows broad spectrum antimicrobial activity.•The LPS binding motif of LG21 has been linked to broad spectrum activity.•LPS binding motif is critical for LG21 bactericidal and membrane activity.•LG21 structure provides mechanistic insight and future antimicrobial drug design.