Divorcing folding from function: how acylation affects the membrane-perturbing properties of an antimicrobial peptide

Brian Stougaard Vad; Line Aagot Hede Thomsen; Kresten Bertelsen; Magnus Franzmann; Jan Mondrup Pedersen; Søren Bang Nielsen; Thomas Vosegaard; Zuzana Valnickova; Troels Skrydstrup; Jan J. Enghild; Reinhard Wimmer; Niels Christian Nielsen; Daniel Otzen

doi:10.1016/j.bbapap.2009.12.006

Divorcing folding from function: how acylation affects the membrane-perturbing properties of an antimicrobial peptide

Brian Stougaard Vad, Line Aagot Hede Thomsen, Kresten Bertelsen, Magnus Franzmann, Jan Mondrup Pedersen, Søren Bang Nielsen, Thomas Vosegaard, Zuzana Valnickova, Troels Skrydstrup, Jan J. Enghild, Reinhard Wimmer, Niels Christian Nielsen, Daniel Otzen

Department of Chemistry - Center for Uopløselige Proteinstrukturer

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

23 Citations (Scopus)

Abstract

Many small cationic peptides, which are unstructured in aqueous solution, have antimicrobial properties. These properties are assumed to be linked to their ability to permeabilize bacterial membranes, accompanied by the transition to an alpha-helical folding state. Here we show that there is no direct link between folding of the antimicrobial peptide Novicidin (Nc) and its membrane permeabilization. N-terminal acylation with C8-C16 alkyl chains and the inclusion of anionic lipids both increase Nc's ability to form alpha-helical structure in the presence of vesicles. Nevertheless, both acylation and anionic lipids reduce the extent of permeabilization of these vesicles and lead to slower permeabilization kinetics. Furthermore, acylation significantly decreases antimicrobial activity. Although acyl chains of increasing length also increase the tendency of the peptides to aggregate in solution, this cannot rationalize our results since permeabilization and antimicrobial activities are observed well below concentrations where aggregation occurs. This suggests that significant induction of alpha-helical structure is not a prerequisite for membrane perturbation in this class of antimicrobial peptides. Our data suggests that for Nc, induction of alpha-helical structure may inhibit rather than facilitate membrane disruption, and that a more peripheral interaction may be the most efficient permeabilization mechanism. Furthermore, acylation leads to a deeper embedding in the membrane, which could lead to an anti-permeabilizing "plugging" effect.

Original language	English
Journal	B B A - Proteins and Proteomics
Volume	1804
Issue	4
Pages (from-to)	806-20
Number of pages	15
ISSN	1570-9639
DOIs	https://doi.org/10.1016/j.bbapap.2009.12.006
Publication status	Published - 2010

Keywords

Acylation
Amino Acid Sequence
Antimicrobial Cationic Peptides
Cell Membrane Permeability
Escherichia coli
Fluorescence Polarization
Liposomes
Membrane Lipids
Micelles
Models, Molecular
Molecular Sequence Data
Nuclear Magnetic Resonance, Biomolecular
Phase Transition
Protein Folding
Protein Multimerization
Protein Structure, Secondary

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10.1016/j.bbapap.2009.12.006

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Vad, B. S., Thomsen, L. A. H., Bertelsen, K., Franzmann, M., Pedersen, J. M., Nielsen, S. B., Vosegaard, T., Valnickova, Z., Skrydstrup, T., Enghild, J. J., Wimmer, R., Nielsen, N. C., & Otzen, D. (2010). Divorcing folding from function: how acylation affects the membrane-perturbing properties of an antimicrobial peptide. B B A - Proteins and Proteomics, 1804(4), 806-20. https://doi.org/10.1016/j.bbapap.2009.12.006

@article{96c339705db311df8b17000ea68e967b,

title = "Divorcing folding from function: how acylation affects the membrane-perturbing properties of an antimicrobial peptide",

abstract = "Many small cationic peptides, which are unstructured in aqueous solution, have antimicrobial properties. These properties are assumed to be linked to their ability to permeabilize bacterial membranes, accompanied by the transition to an alpha-helical folding state. Here we show that there is no direct link between folding of the antimicrobial peptide Novicidin (Nc) and its membrane permeabilization. N-terminal acylation with C8-C16 alkyl chains and the inclusion of anionic lipids both increase Nc's ability to form alpha-helical structure in the presence of vesicles. Nevertheless, both acylation and anionic lipids reduce the extent of permeabilization of these vesicles and lead to slower permeabilization kinetics. Furthermore, acylation significantly decreases antimicrobial activity. Although acyl chains of increasing length also increase the tendency of the peptides to aggregate in solution, this cannot rationalize our results since permeabilization and antimicrobial activities are observed well below concentrations where aggregation occurs. This suggests that significant induction of alpha-helical structure is not a prerequisite for membrane perturbation in this class of antimicrobial peptides. Our data suggests that for Nc, induction of alpha-helical structure may inhibit rather than facilitate membrane disruption, and that a more peripheral interaction may be the most efficient permeabilization mechanism. Furthermore, acylation leads to a deeper embedding in the membrane, which could lead to an anti-permeabilizing {"}plugging{"} effect.",

keywords = "Acylation, Amino Acid Sequence, Antimicrobial Cationic Peptides, Cell Membrane Permeability, Escherichia coli, Fluorescence Polarization, Liposomes, Membrane Lipids, Micelles, Models, Molecular, Molecular Sequence Data, Nuclear Magnetic Resonance, Biomolecular, Phase Transition, Protein Folding, Protein Multimerization, Protein Structure, Secondary",

author = "Vad, {Brian Stougaard} and Thomsen, {Line Aagot Hede} and Kresten Bertelsen and Magnus Franzmann and Pedersen, {Jan Mondrup} and Nielsen, {S{\o}ren Bang} and Thomas Vosegaard and Zuzana Valnickova and Troels Skrydstrup and Enghild, {Jan J.} and Reinhard Wimmer and Nielsen, {Niels Christian} and Daniel Otzen",

year = "2010",

doi = "10.1016/j.bbapap.2009.12.006",

language = "English",

volume = "1804",

pages = "806--20",

journal = "B B A - Proteins and Proteomics",

issn = "1570-9639",

publisher = "Elsevier B.V.",

number = "4",

}

Vad, BS, Thomsen, LAH, Bertelsen, K, Franzmann, M, Pedersen, JM, Nielsen, SB, Vosegaard, T, Valnickova, Z, Skrydstrup, T , Enghild, JJ, Wimmer, R, Nielsen, NC & Otzen, D 2010, 'Divorcing folding from function: how acylation affects the membrane-perturbing properties of an antimicrobial peptide', B B A - Proteins and Proteomics, vol. 1804, no. 4, pp. 806-20. https://doi.org/10.1016/j.bbapap.2009.12.006

Divorcing folding from function: how acylation affects the membrane-perturbing properties of an antimicrobial peptide. / Vad, Brian Stougaard; Thomsen, Line Aagot Hede; Bertelsen, Kresten et al.
In: B B A - Proteins and Proteomics, Vol. 1804, No. 4, 2010, p. 806-20.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

TY - JOUR

T1 - Divorcing folding from function: how acylation affects the membrane-perturbing properties of an antimicrobial peptide

AU - Vad, Brian Stougaard

AU - Thomsen, Line Aagot Hede

AU - Bertelsen, Kresten

AU - Franzmann, Magnus

AU - Pedersen, Jan Mondrup

AU - Nielsen, Søren Bang

AU - Vosegaard, Thomas

AU - Valnickova, Zuzana

AU - Skrydstrup, Troels

AU - Enghild, Jan J.

AU - Wimmer, Reinhard

AU - Nielsen, Niels Christian

AU - Otzen, Daniel

PY - 2010

Y1 - 2010

N2 - Many small cationic peptides, which are unstructured in aqueous solution, have antimicrobial properties. These properties are assumed to be linked to their ability to permeabilize bacterial membranes, accompanied by the transition to an alpha-helical folding state. Here we show that there is no direct link between folding of the antimicrobial peptide Novicidin (Nc) and its membrane permeabilization. N-terminal acylation with C8-C16 alkyl chains and the inclusion of anionic lipids both increase Nc's ability to form alpha-helical structure in the presence of vesicles. Nevertheless, both acylation and anionic lipids reduce the extent of permeabilization of these vesicles and lead to slower permeabilization kinetics. Furthermore, acylation significantly decreases antimicrobial activity. Although acyl chains of increasing length also increase the tendency of the peptides to aggregate in solution, this cannot rationalize our results since permeabilization and antimicrobial activities are observed well below concentrations where aggregation occurs. This suggests that significant induction of alpha-helical structure is not a prerequisite for membrane perturbation in this class of antimicrobial peptides. Our data suggests that for Nc, induction of alpha-helical structure may inhibit rather than facilitate membrane disruption, and that a more peripheral interaction may be the most efficient permeabilization mechanism. Furthermore, acylation leads to a deeper embedding in the membrane, which could lead to an anti-permeabilizing "plugging" effect.

AB - Many small cationic peptides, which are unstructured in aqueous solution, have antimicrobial properties. These properties are assumed to be linked to their ability to permeabilize bacterial membranes, accompanied by the transition to an alpha-helical folding state. Here we show that there is no direct link between folding of the antimicrobial peptide Novicidin (Nc) and its membrane permeabilization. N-terminal acylation with C8-C16 alkyl chains and the inclusion of anionic lipids both increase Nc's ability to form alpha-helical structure in the presence of vesicles. Nevertheless, both acylation and anionic lipids reduce the extent of permeabilization of these vesicles and lead to slower permeabilization kinetics. Furthermore, acylation significantly decreases antimicrobial activity. Although acyl chains of increasing length also increase the tendency of the peptides to aggregate in solution, this cannot rationalize our results since permeabilization and antimicrobial activities are observed well below concentrations where aggregation occurs. This suggests that significant induction of alpha-helical structure is not a prerequisite for membrane perturbation in this class of antimicrobial peptides. Our data suggests that for Nc, induction of alpha-helical structure may inhibit rather than facilitate membrane disruption, and that a more peripheral interaction may be the most efficient permeabilization mechanism. Furthermore, acylation leads to a deeper embedding in the membrane, which could lead to an anti-permeabilizing "plugging" effect.

KW - Acylation

KW - Amino Acid Sequence

KW - Antimicrobial Cationic Peptides

KW - Cell Membrane Permeability

KW - Escherichia coli

KW - Fluorescence Polarization

KW - Liposomes

KW - Membrane Lipids

KW - Micelles

KW - Models, Molecular

KW - Molecular Sequence Data

KW - Nuclear Magnetic Resonance, Biomolecular

KW - Phase Transition

KW - Protein Folding

KW - Protein Multimerization

KW - Protein Structure, Secondary

U2 - 10.1016/j.bbapap.2009.12.006

DO - 10.1016/j.bbapap.2009.12.006

M3 - Journal article

C2 - 20026432

SN - 1570-9639

VL - 1804

SP - 806

EP - 820

JO - B B A - Proteins and Proteomics

JF - B B A - Proteins and Proteomics

IS - 4

ER -

Divorcing folding from function: how acylation affects the membrane-perturbing properties of an antimicrobial peptide

Abstract

Keywords

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