title: The role of C-terminal amidation in the membrane interactions of the anionic antimicrobial peptide, maximin H5.
abstract: Maximin H5 is an anionic antimicrobial peptide from amphibians, which carries a C-terminal amide moiety, and was found to be moderately haemolytic (20%). The α-helicity of the peptide was 42% in the presence of lipid mimics of erythrocyte membranes and was found able to penetrate (10.8 mN m(-1)) and lyse these model membranes (64 %). In contrast, the deaminated peptide exhibited lower levels of haemolysis (12%) and α-helicity (16%) along with a reduced ability to penetrate (7.8 m Nm(-1)) and lyse (55%) lipid mimics of erythrocyte membranes. Taken with molecular dynamic simulations and theoretical analysis, these data suggest that native maximin H5 primarily exerts its haemolytic action via the formation of an oblique orientated α-helical structure and tilted membrane insertion. However, the C-terminal deamination of maximin H5 induces a loss of tilted α-helical structure, which abolishes the ability of the peptide's N-terminal and C-terminal regions to H-bond and leads to a loss in haemolytic ability. Taken in combination, these observations strongly suggest that the C-terminal amide moiety carried by maximin H5 is required to stabilise the adoption of membrane interactive tilted structure by the peptide. Consistent with previous reports, these data show that the efficacy of interaction and specificity of maximin H5 for membranes can be attenuated by sequence modification and may assist in the development of variants of the peptide with the potential to serve as anti-infectives.
authors: School of Pharmacy and Biomedical Sciences, University of Central Lancashire, Preston PR1 2HE, UK; School of Applied Science, London South Bank University, 103 Borough Road, London SE1 0AA, UK.
authors: Sarah R
authors: SR
authors: Dennison
authors: School of Computing Engineering and Physical Science, University of Central Lancashire, Preston PR1 2HE, UK.
authors: Manuela
authors: M
authors: Mura
authors: School of Applied Science, London South Bank University, 103 Borough Road, London SE1 0AA, UK; School of Forensic and Investigative Science, University of Central Lancashire, Preston PR1 2HE, UK.
authors: Frederick
authors: F
authors: Harris
authors: School of Forensic and Investigative Science, University of Central Lancashire, Preston PR1 2HE, UK.
authors: Leslie H G
authors: LH
authors: Morton
authors: School of Mathematics and Physics, University of Lincoln, Lincolnshire LN6 7TS, UK.
authors: Andrei
authors: A
authors: Zvelindovsky
authors: School of Applied Science, London South Bank University, 103 Borough Road, London SE1 0AA, UK; School of Forensic and Investigative Science, University of Central Lancashire, Preston PR1 2HE, UK. Electronic address: phoenixd@lsbu.ac.uk.
authors: David A
authors: DA
authors: Phoenix
mesh_terms: D000577:Amides / Q000737:chemistry / Q000494:pharmacology* / Q000633:toxicity; D029845:Amphibian Proteins / Q000737:chemistry / Q000494:pharmacology* / Q000633:toxicity; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology* / Q000633:toxicity; D002462:Cell Membrane / Q000187:drug effects*; D002463:Cell Membrane Permeability / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004910:Erythrocyte Membrane / Q000187:drug effects*; D004926:Escherichia coli / Q000187:drug effects* / Q000254:growth & development; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D008563:Membrane Lipids / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D056004:Molecular Dynamics Simulation; D010446:Peptide Fragments / Q000737:chemistry / Q000494:pharmacology* / Q000633:toxicity; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013211:Staphylococcus aureus / Q000187:drug effects* / Q000254:growth & development; D013329:Structure-Activity Relationship; D013997:Time Factors
chemical_list: D000577:Amides; D029845:Amphibian Proteins; D000900:Anti-Bacterial Agents; D008563:Membrane Lipids; D010446:Peptide Fragments; C464648:maximin H5, Bombina maxima
keywords: Anionic antimicrobial peptide; Haemolysis; Lipid monolayer; Lysis; α-Helical