Identification Of A Novel Histone Derived Antimicrobial Peptide In Airway Surface Liquid.
The airway surface liquid (ASL), a protective layer secreted by the airway epithelium, represents the first line of defence against inhaled infectious material. It contains a complex array of secreted proteins and peptides that aid the neutralisation and removal of inhaled microbes and toxicants. Th...
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crwiley:10.1096/fasebj.2020.34.s1.02889 2024-06-02T08:03:43+00:00 Identification Of A Novel Histone Derived Antimicrobial Peptide In Airway Surface Liquid. Biggart, Matthew Guy Stevenson Ling, Xie Simpson, Nathan Gani, Jurnorain Wrobel, John Hilpert, Kai Chen, Xian Tarran, Robert Baines, Deborah 2020 http://dx.doi.org/10.1096/fasebj.2020.34.s1.02889 en eng Wiley http://onlinelibrary.wiley.com/termsAndConditions#vor The FASEB Journal volume 34, issue S1, page 1-1 ISSN 0892-6638 1530-6860 journal-article 2020 crwiley https://doi.org/10.1096/fasebj.2020.34.s1.02889 2024-05-03T11:08:29Z The airway surface liquid (ASL), a protective layer secreted by the airway epithelium, represents the first line of defence against inhaled infectious material. It contains a complex array of secreted proteins and peptides that aid the neutralisation and removal of inhaled microbes and toxicants. The ASL also contains many proteases that can cleave proteins to generate further bioactive peptides. We therefore hypothesised that the airway ASL sustained an extensive peptidome containing novel bioactive peptides. We examined the ASL peptide profile of normal human bronchial epithelial cells transformed with BMI‐1 (NHBE‐BMI1) and cell‐lines Calu3 (submucosal adenocarcinoma) and H441 (Clara cell like adenocarcinoma). ASL was acquired by washing the apical surface of epithelial monolayers grown at air‐liquid interface with PBS (100μl) at 0, 24 and 120 hours after exposure to hyperglycaemic (25mM glucose) or normoglycaemic (5mM glucose and 20mM mannitol) basolateral medium. The ASL peptides < 10kDa, were subsequently isolated and analysed using the Q Exactive™ HF‐X Hybrid Quadrupole‐Orbitrap™ Mass Spectrometer. The resulting spectra of native peptides and their subsequent fragments were analysed using the pNovo De novo sequencing tool. Our preliminary results identified 4765 unique peptides (NHBE‐BMI1: 646, Calu3: 3150, H441: 1197). Of these, 37 peptides were common to all samples regardless of glycaemic state. A number of histone derived peptides were identified in all samples and cell lines. These peptides had a proline‐alanine rich N‐terminal region and a cationic C‐terminal, similar to antimicrobial peptides from histone H1 found in Atlantic salmon skin mucous. Synthetic peptides were produced by SPOT synthesis and the antimicrobial activity tested against luminescent Pseudomonas aeruginosa over 18 hours at 37°C in (100mM Tris (pH 7) and 20mM glucose). One of the crude peptides elicited a dose dependent inhibition of bacterial luminescence at 4.63±1.34 μM (n=3), indicating no bacteria survived. Truncated ... Article in Journal/Newspaper Atlantic salmon Wiley Online Library The FASEB Journal 34 S1 1 1 |
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Wiley Online Library |
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English |
description |
The airway surface liquid (ASL), a protective layer secreted by the airway epithelium, represents the first line of defence against inhaled infectious material. It contains a complex array of secreted proteins and peptides that aid the neutralisation and removal of inhaled microbes and toxicants. The ASL also contains many proteases that can cleave proteins to generate further bioactive peptides. We therefore hypothesised that the airway ASL sustained an extensive peptidome containing novel bioactive peptides. We examined the ASL peptide profile of normal human bronchial epithelial cells transformed with BMI‐1 (NHBE‐BMI1) and cell‐lines Calu3 (submucosal adenocarcinoma) and H441 (Clara cell like adenocarcinoma). ASL was acquired by washing the apical surface of epithelial monolayers grown at air‐liquid interface with PBS (100μl) at 0, 24 and 120 hours after exposure to hyperglycaemic (25mM glucose) or normoglycaemic (5mM glucose and 20mM mannitol) basolateral medium. The ASL peptides < 10kDa, were subsequently isolated and analysed using the Q Exactive™ HF‐X Hybrid Quadrupole‐Orbitrap™ Mass Spectrometer. The resulting spectra of native peptides and their subsequent fragments were analysed using the pNovo De novo sequencing tool. Our preliminary results identified 4765 unique peptides (NHBE‐BMI1: 646, Calu3: 3150, H441: 1197). Of these, 37 peptides were common to all samples regardless of glycaemic state. A number of histone derived peptides were identified in all samples and cell lines. These peptides had a proline‐alanine rich N‐terminal region and a cationic C‐terminal, similar to antimicrobial peptides from histone H1 found in Atlantic salmon skin mucous. Synthetic peptides were produced by SPOT synthesis and the antimicrobial activity tested against luminescent Pseudomonas aeruginosa over 18 hours at 37°C in (100mM Tris (pH 7) and 20mM glucose). One of the crude peptides elicited a dose dependent inhibition of bacterial luminescence at 4.63±1.34 μM (n=3), indicating no bacteria survived. Truncated ... |
format |
Article in Journal/Newspaper |
author |
Biggart, Matthew Guy Stevenson Ling, Xie Simpson, Nathan Gani, Jurnorain Wrobel, John Hilpert, Kai Chen, Xian Tarran, Robert Baines, Deborah |
spellingShingle |
Biggart, Matthew Guy Stevenson Ling, Xie Simpson, Nathan Gani, Jurnorain Wrobel, John Hilpert, Kai Chen, Xian Tarran, Robert Baines, Deborah Identification Of A Novel Histone Derived Antimicrobial Peptide In Airway Surface Liquid. |
author_facet |
Biggart, Matthew Guy Stevenson Ling, Xie Simpson, Nathan Gani, Jurnorain Wrobel, John Hilpert, Kai Chen, Xian Tarran, Robert Baines, Deborah |
author_sort |
Biggart, Matthew Guy Stevenson |
title |
Identification Of A Novel Histone Derived Antimicrobial Peptide In Airway Surface Liquid. |
title_short |
Identification Of A Novel Histone Derived Antimicrobial Peptide In Airway Surface Liquid. |
title_full |
Identification Of A Novel Histone Derived Antimicrobial Peptide In Airway Surface Liquid. |
title_fullStr |
Identification Of A Novel Histone Derived Antimicrobial Peptide In Airway Surface Liquid. |
title_full_unstemmed |
Identification Of A Novel Histone Derived Antimicrobial Peptide In Airway Surface Liquid. |
title_sort |
identification of a novel histone derived antimicrobial peptide in airway surface liquid. |
publisher |
Wiley |
publishDate |
2020 |
url |
http://dx.doi.org/10.1096/fasebj.2020.34.s1.02889 |
genre |
Atlantic salmon |
genre_facet |
Atlantic salmon |
op_source |
The FASEB Journal volume 34, issue S1, page 1-1 ISSN 0892-6638 1530-6860 |
op_rights |
http://onlinelibrary.wiley.com/termsAndConditions#vor |
op_doi |
https://doi.org/10.1096/fasebj.2020.34.s1.02889 |
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The FASEB Journal |
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34 |
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S1 |
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1 |
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1 |
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1800748327337197568 |