Structural basis for endotoxin neutralisation and anti-inflammatory activity of thrombin-derived C-terminal peptides

Rathi Saravanan; Daniel A Holdbrook; Jitka Petrlova; Shalini Singh; Nils A Berglund; Yeu Khai Choong; Sven Kjellström; Peter J Bond; Martin Malmsten; Artur Schmidtchen

doi:10.1038/s41467-018-05242-0

Structural basis for endotoxin neutralisation and anti-inflammatory activity of thrombin-derived C-terminal peptides

Rathi Saravanan, Daniel A Holdbrook, Jitka Petrlova, Shalini Singh, Nils A Berglund, Yeu Khai Choong, Sven Kjellström, Peter J Bond, Martin Malmsten, Artur Schmidtchen

Dermatology, Department of

Abstract

Thrombin-derived C-terminal peptides (TCPs) of about 2 kDa are present in wounds, where they exert anti-endotoxic functions. Employing a combination of nuclear magnetic resonance spectroscopy (NMR), biophysical, mass spectrometry and cellular studies combined with in silico multiscale modelling, we here determine the bound conformation of HVF18 (HVFRLKKWIQKVIDQFGE), a TCP generated by neutrophil elastase, in complex with bacterial lipopolysaccharide (LPS) and define a previously undisclosed interaction between TCPs and human CD14. Further, we show that TCPs bind to the LPS-binding hydrophobic pocket of CD14 and identify the peptide region crucial for TCP interaction with LPS and CD14. Taken together, our results demonstrate the role of structural transitions in LPS complex formation and CD14 interaction, providing a molecular explanation for the previously observed therapeutic effects of TCPs in experimental models of bacterial sepsis and endotoxin shock.

Original language	English
Journal	Nature Communications
Volume	9
Issue number	1
Pages (from-to)	2762
ISSN	2041-1722
DOIs	https://doi.org/10.1038/s41467-018-05242-0
Publication status	Published - 17 Jul 2018

Keywords

Amino Acid Sequence
Antimicrobial Cationic Peptides/chemistry
Binding Sites
Escherichia coli/genetics
Humans
Hydrophobic and Hydrophilic Interactions
Leukocyte Elastase/chemistry
Lipopolysaccharide Receptors/chemistry
Lipopolysaccharides/chemistry
Neutralization Tests
Protein Binding
Protein Conformation, alpha-Helical
Protein Conformation, beta-Strand
Protein Interaction Domains and Motifs
THP-1 Cells
Thrombin/chemistry

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title = "Structural basis for endotoxin neutralisation and anti-inflammatory activity of thrombin-derived C-terminal peptides",

abstract = "Thrombin-derived C-terminal peptides (TCPs) of about 2 kDa are present in wounds, where they exert anti-endotoxic functions. Employing a combination of nuclear magnetic resonance spectroscopy (NMR), biophysical, mass spectrometry and cellular studies combined with in silico multiscale modelling, we here determine the bound conformation of HVF18 (HVFRLKKWIQKVIDQFGE), a TCP generated by neutrophil elastase, in complex with bacterial lipopolysaccharide (LPS) and define a previously undisclosed interaction between TCPs and human CD14. Further, we show that TCPs bind to the LPS-binding hydrophobic pocket of CD14 and identify the peptide region crucial for TCP interaction with LPS and CD14. Taken together, our results demonstrate the role of structural transitions in LPS complex formation and CD14 interaction, providing a molecular explanation for the previously observed therapeutic effects of TCPs in experimental models of bacterial sepsis and endotoxin shock.",

keywords = "Amino Acid Sequence, Antimicrobial Cationic Peptides/chemistry, Binding Sites, Escherichia coli/genetics, Humans, Hydrophobic and Hydrophilic Interactions, Leukocyte Elastase/chemistry, Lipopolysaccharide Receptors/chemistry, Lipopolysaccharides/chemistry, Neutralization Tests, Protein Binding, Protein Conformation, alpha-Helical, Protein Conformation, beta-Strand, Protein Interaction Domains and Motifs, THP-1 Cells, Thrombin/chemistry",

author = "Rathi Saravanan and Holdbrook, {Daniel A} and Jitka Petrlova and Shalini Singh and Berglund, {Nils A} and Choong, {Yeu Khai} and Sven Kjellstr{\"o}m and Bond, {Peter J} and Martin Malmsten and Artur Schmidtchen",

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doi = "10.1038/s41467-018-05242-0",

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T1 - Structural basis for endotoxin neutralisation and anti-inflammatory activity of thrombin-derived C-terminal peptides

AU - Saravanan, Rathi

AU - Holdbrook, Daniel A

AU - Petrlova, Jitka

AU - Singh, Shalini

AU - Berglund, Nils A

AU - Choong, Yeu Khai

AU - Kjellström, Sven

AU - Bond, Peter J

AU - Malmsten, Martin

AU - Schmidtchen, Artur

PY - 2018/7/17

Y1 - 2018/7/17

N2 - Thrombin-derived C-terminal peptides (TCPs) of about 2 kDa are present in wounds, where they exert anti-endotoxic functions. Employing a combination of nuclear magnetic resonance spectroscopy (NMR), biophysical, mass spectrometry and cellular studies combined with in silico multiscale modelling, we here determine the bound conformation of HVF18 (HVFRLKKWIQKVIDQFGE), a TCP generated by neutrophil elastase, in complex with bacterial lipopolysaccharide (LPS) and define a previously undisclosed interaction between TCPs and human CD14. Further, we show that TCPs bind to the LPS-binding hydrophobic pocket of CD14 and identify the peptide region crucial for TCP interaction with LPS and CD14. Taken together, our results demonstrate the role of structural transitions in LPS complex formation and CD14 interaction, providing a molecular explanation for the previously observed therapeutic effects of TCPs in experimental models of bacterial sepsis and endotoxin shock.

AB - Thrombin-derived C-terminal peptides (TCPs) of about 2 kDa are present in wounds, where they exert anti-endotoxic functions. Employing a combination of nuclear magnetic resonance spectroscopy (NMR), biophysical, mass spectrometry and cellular studies combined with in silico multiscale modelling, we here determine the bound conformation of HVF18 (HVFRLKKWIQKVIDQFGE), a TCP generated by neutrophil elastase, in complex with bacterial lipopolysaccharide (LPS) and define a previously undisclosed interaction between TCPs and human CD14. Further, we show that TCPs bind to the LPS-binding hydrophobic pocket of CD14 and identify the peptide region crucial for TCP interaction with LPS and CD14. Taken together, our results demonstrate the role of structural transitions in LPS complex formation and CD14 interaction, providing a molecular explanation for the previously observed therapeutic effects of TCPs in experimental models of bacterial sepsis and endotoxin shock.

KW - Amino Acid Sequence

KW - Antimicrobial Cationic Peptides/chemistry

KW - Binding Sites

KW - Escherichia coli/genetics

KW - Humans

KW - Hydrophobic and Hydrophilic Interactions

KW - Leukocyte Elastase/chemistry

KW - Lipopolysaccharide Receptors/chemistry

KW - Lipopolysaccharides/chemistry

KW - Neutralization Tests

KW - Protein Binding

KW - Protein Conformation, alpha-Helical

KW - Protein Conformation, beta-Strand

KW - Protein Interaction Domains and Motifs

KW - THP-1 Cells

KW - Thrombin/chemistry

U2 - 10.1038/s41467-018-05242-0

DO - 10.1038/s41467-018-05242-0

M3 - Journal article

C2 - 30018388

VL - 9

SP - 2762

JO - Nature Communications

JF - Nature Communications

SN - 2041-1722

IS - 1

ER -

Structural basis for endotoxin neutralisation and anti-inflammatory activity of thrombin-derived C-terminal peptides

Abstract

Keywords

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