Structure, activity and interactions of the cysteine deleted analog of tachyplesin-1 with lipopolysaccharide micelle : mechanistic insights into outer-membrane permeabilization and endotoxin neutralization

Tachyplesin-1, a disulfide stabilized β-hairpin antimicrobial peptide, can be found at the hemocytes of horse shoe crab Tachypleus tridentatus. A cysteine deleted linear analog of tachyplesin-1 or CDT (KWFRVYRGIYRRR-NH2) contains a broad spectrum of bactericidal activity with a reduced hemolytic pro...

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Main Authors: Saravanan, Rathi, Mohanram, Harini, Joshi, Mangesh, Torres, Jaume, Ruedl, Christiane, Bhattacharjya, Surajit, Domadia, Prerna N.
Other Authors: School of Biological Sciences
Format: Article
Language:English
Published: 2013
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Online Access:https://hdl.handle.net/10356/98894
http://hdl.handle.net/10220/12689
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Institution: Nanyang Technological University
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spelling sg-ntu-dr.10356-988942020-03-07T12:18:19Z Structure, activity and interactions of the cysteine deleted analog of tachyplesin-1 with lipopolysaccharide micelle : mechanistic insights into outer-membrane permeabilization and endotoxin neutralization Saravanan, Rathi Mohanram, Harini Joshi, Mangesh Torres, Jaume Ruedl, Christiane Bhattacharjya, Surajit Domadia, Prerna N. School of Biological Sciences DRNTU::Science::Biological sciences Tachyplesin-1, a disulfide stabilized β-hairpin antimicrobial peptide, can be found at the hemocytes of horse shoe crab Tachypleus tridentatus. A cysteine deleted linear analog of tachyplesin-1 or CDT (KWFRVYRGIYRRR-NH2) contains a broad spectrum of bactericidal activity with a reduced hemolytic property. The bactericidal activity of CDT stems from selective interactions with the negatively charged lipids including LPS. In this work, CDT–LPS interactions were investigated using NMR spectroscopy, optical spectroscopy and functional assays. We found that CDT neutralized LPS and disrupted permeability barrier of the outer membrane. Zeta potential and ITC studies demonstrated charge compensation and hydrophobic interactions of CDT with the LPS-outer membrane, respectively. Secondary structure of the peptide was probed by CD and FT-IR experiments indicating β-strands and/or β-turn conformations in the LPS micelle. An ensemble of structures, determined in LPS micelle by NMR, revealed a β-hairpin like topology of the CDT peptide that was typified by an extended cationic surface and a relatively shorter segment of hydrophobic region. Interestingly, at the non-polar face, residue R11 was found to be in a close proximity to the indole ring of W2, suggesting a cation–π type interactions. Further, saturation transfer difference (STD) NMR studies established intimate contacts among the aromatic and cationic residues of CDT with the LPS micelle. Fluorescence and dynamic light scattering experiments demonstrated that CDT imparted structural destabilization to the aggregated states of LPS. Collectively, atomic resolution structure and interactions of CDT with the outer membrane-LPS could be exploited for developing potent broad spectrum antimicrobial and anti-sepsis agents. 2013-08-01T01:41:29Z 2019-12-06T20:00:52Z 2013-08-01T01:41:29Z 2019-12-06T20:00:52Z 2012 2012 Journal Article Saravanan, R., Mohanram, H., Joshi, M., Domadia, P. N., Torres, J., Ruedl, C.,& Bhattacharjya, S. (2012). Structure, activity and interactions of the cysteine deleted analog of tachyplesin-1 with lipopolysaccharide micelle: Mechanistic insights into outer-membrane permeabilization and endotoxin neutralization. Biochimica et Biophysica Acta (BBA) - Biomembranes, 1818(7), 1613-1624. 0005-2736 https://hdl.handle.net/10356/98894 http://hdl.handle.net/10220/12689 10.1016/j.bbamem.2012.03.015 en Biochimica et Biophysica Acta (BBA) - biomembranes
institution Nanyang Technological University
building NTU Library
country Singapore
collection DR-NTU
language English
topic DRNTU::Science::Biological sciences
spellingShingle DRNTU::Science::Biological sciences
Saravanan, Rathi
Mohanram, Harini
Joshi, Mangesh
Torres, Jaume
Ruedl, Christiane
Bhattacharjya, Surajit
Domadia, Prerna N.
Structure, activity and interactions of the cysteine deleted analog of tachyplesin-1 with lipopolysaccharide micelle : mechanistic insights into outer-membrane permeabilization and endotoxin neutralization
description Tachyplesin-1, a disulfide stabilized β-hairpin antimicrobial peptide, can be found at the hemocytes of horse shoe crab Tachypleus tridentatus. A cysteine deleted linear analog of tachyplesin-1 or CDT (KWFRVYRGIYRRR-NH2) contains a broad spectrum of bactericidal activity with a reduced hemolytic property. The bactericidal activity of CDT stems from selective interactions with the negatively charged lipids including LPS. In this work, CDT–LPS interactions were investigated using NMR spectroscopy, optical spectroscopy and functional assays. We found that CDT neutralized LPS and disrupted permeability barrier of the outer membrane. Zeta potential and ITC studies demonstrated charge compensation and hydrophobic interactions of CDT with the LPS-outer membrane, respectively. Secondary structure of the peptide was probed by CD and FT-IR experiments indicating β-strands and/or β-turn conformations in the LPS micelle. An ensemble of structures, determined in LPS micelle by NMR, revealed a β-hairpin like topology of the CDT peptide that was typified by an extended cationic surface and a relatively shorter segment of hydrophobic region. Interestingly, at the non-polar face, residue R11 was found to be in a close proximity to the indole ring of W2, suggesting a cation–π type interactions. Further, saturation transfer difference (STD) NMR studies established intimate contacts among the aromatic and cationic residues of CDT with the LPS micelle. Fluorescence and dynamic light scattering experiments demonstrated that CDT imparted structural destabilization to the aggregated states of LPS. Collectively, atomic resolution structure and interactions of CDT with the outer membrane-LPS could be exploited for developing potent broad spectrum antimicrobial and anti-sepsis agents.
author2 School of Biological Sciences
author_facet School of Biological Sciences
Saravanan, Rathi
Mohanram, Harini
Joshi, Mangesh
Torres, Jaume
Ruedl, Christiane
Bhattacharjya, Surajit
Domadia, Prerna N.
format Article
author Saravanan, Rathi
Mohanram, Harini
Joshi, Mangesh
Torres, Jaume
Ruedl, Christiane
Bhattacharjya, Surajit
Domadia, Prerna N.
author_sort Saravanan, Rathi
title Structure, activity and interactions of the cysteine deleted analog of tachyplesin-1 with lipopolysaccharide micelle : mechanistic insights into outer-membrane permeabilization and endotoxin neutralization
title_short Structure, activity and interactions of the cysteine deleted analog of tachyplesin-1 with lipopolysaccharide micelle : mechanistic insights into outer-membrane permeabilization and endotoxin neutralization
title_full Structure, activity and interactions of the cysteine deleted analog of tachyplesin-1 with lipopolysaccharide micelle : mechanistic insights into outer-membrane permeabilization and endotoxin neutralization
title_fullStr Structure, activity and interactions of the cysteine deleted analog of tachyplesin-1 with lipopolysaccharide micelle : mechanistic insights into outer-membrane permeabilization and endotoxin neutralization
title_full_unstemmed Structure, activity and interactions of the cysteine deleted analog of tachyplesin-1 with lipopolysaccharide micelle : mechanistic insights into outer-membrane permeabilization and endotoxin neutralization
title_sort structure, activity and interactions of the cysteine deleted analog of tachyplesin-1 with lipopolysaccharide micelle : mechanistic insights into outer-membrane permeabilization and endotoxin neutralization
publishDate 2013
url https://hdl.handle.net/10356/98894
http://hdl.handle.net/10220/12689
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