Multi-Approach Analysis for the Identification of Proteases within Birch Pollen

Birch pollen allergy is highly prevalent, with up to 100 million reported cases worldwide. Proteases in such allergen sources have been suggested to contribute to primary sensitisation and exacerbation of allergic disorders. Until now the protease content of Betula verrucosa, a birch species endemic...

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Main Authors: Olivia E. McKenna, Gernot Posselt, Peter Briza, Peter Lackner, Armin O. Schmitt, Gabriele Gadermaier, Silja Wessler, Fatima Ferreira
Format: Article
Language:English
Published: MDPI AG 2017-07-01
Series:International Journal of Molecular Sciences
Subjects:
Online Access:https://www.mdpi.com/1422-0067/18/7/1433
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spelling doaj-1062b96843964997afdd32b10e34e11b2020-11-24T23:14:52ZengMDPI AGInternational Journal of Molecular Sciences1422-00672017-07-01187143310.3390/ijms18071433ijms18071433Multi-Approach Analysis for the Identification of Proteases within Birch PollenOlivia E. McKenna0Gernot Posselt1Peter Briza2Peter Lackner3Armin O. Schmitt4Gabriele Gadermaier5Silja Wessler6Fatima Ferreira7Department of Molecular Biology, University of Salzburg, Salzburg 5020, AustriaDepartment of Molecular Biology, University of Salzburg, Salzburg 5020, AustriaDepartment of Molecular Biology, University of Salzburg, Salzburg 5020, AustriaDepartment of Molecular Biology, University of Salzburg, Salzburg 5020, AustriaDepartment of Breeding Informatics, Georg August-Universität Göttingen, Göttingen 37073, GermanyDepartment of Molecular Biology, University of Salzburg, Salzburg 5020, AustriaDepartment of Molecular Biology, University of Salzburg, Salzburg 5020, AustriaDepartment of Molecular Biology, University of Salzburg, Salzburg 5020, AustriaBirch pollen allergy is highly prevalent, with up to 100 million reported cases worldwide. Proteases in such allergen sources have been suggested to contribute to primary sensitisation and exacerbation of allergic disorders. Until now the protease content of Betula verrucosa, a birch species endemic to the northern hemisphere has not been studied in detail. Hence, we aim to identify and characterise pollen and bacteria-derived proteases found within birch pollen. The pollen transcriptome was constructed via de novo transcriptome sequencing and analysis of the proteome was achieved via mass spectrometry; a cross-comparison of the two databases was then performed. A total of 42 individual proteases were identified at the proteomic level. Further clustering of proteases into their distinct catalytic classes revealed serine, cysteine, aspartic, threonine, and metallo-proteases. Further to this, protease activity of the pollen was quantified using a fluorescently-labelled casein substrate protease assay, as 0.61 ng/mg of pollen. A large number of bacterial strains were isolated from freshly collected birch pollen and zymographic gels with gelatinase and casein, enabled visualisation of proteolytic activity of the pollen and the collected bacterial strains. We report the successful discovery of pollen and bacteria-derived proteases of Betula verrucosa.https://www.mdpi.com/1422-0067/18/7/1433birch pollenallergyproteaseproteometranscriptomezymogram
collection DOAJ
language English
format Article
sources DOAJ
author Olivia E. McKenna
Gernot Posselt
Peter Briza
Peter Lackner
Armin O. Schmitt
Gabriele Gadermaier
Silja Wessler
Fatima Ferreira
spellingShingle Olivia E. McKenna
Gernot Posselt
Peter Briza
Peter Lackner
Armin O. Schmitt
Gabriele Gadermaier
Silja Wessler
Fatima Ferreira
Multi-Approach Analysis for the Identification of Proteases within Birch Pollen
International Journal of Molecular Sciences
birch pollen
allergy
protease
proteome
transcriptome
zymogram
author_facet Olivia E. McKenna
Gernot Posselt
Peter Briza
Peter Lackner
Armin O. Schmitt
Gabriele Gadermaier
Silja Wessler
Fatima Ferreira
author_sort Olivia E. McKenna
title Multi-Approach Analysis for the Identification of Proteases within Birch Pollen
title_short Multi-Approach Analysis for the Identification of Proteases within Birch Pollen
title_full Multi-Approach Analysis for the Identification of Proteases within Birch Pollen
title_fullStr Multi-Approach Analysis for the Identification of Proteases within Birch Pollen
title_full_unstemmed Multi-Approach Analysis for the Identification of Proteases within Birch Pollen
title_sort multi-approach analysis for the identification of proteases within birch pollen
publisher MDPI AG
series International Journal of Molecular Sciences
issn 1422-0067
publishDate 2017-07-01
description Birch pollen allergy is highly prevalent, with up to 100 million reported cases worldwide. Proteases in such allergen sources have been suggested to contribute to primary sensitisation and exacerbation of allergic disorders. Until now the protease content of Betula verrucosa, a birch species endemic to the northern hemisphere has not been studied in detail. Hence, we aim to identify and characterise pollen and bacteria-derived proteases found within birch pollen. The pollen transcriptome was constructed via de novo transcriptome sequencing and analysis of the proteome was achieved via mass spectrometry; a cross-comparison of the two databases was then performed. A total of 42 individual proteases were identified at the proteomic level. Further clustering of proteases into their distinct catalytic classes revealed serine, cysteine, aspartic, threonine, and metallo-proteases. Further to this, protease activity of the pollen was quantified using a fluorescently-labelled casein substrate protease assay, as 0.61 ng/mg of pollen. A large number of bacterial strains were isolated from freshly collected birch pollen and zymographic gels with gelatinase and casein, enabled visualisation of proteolytic activity of the pollen and the collected bacterial strains. We report the successful discovery of pollen and bacteria-derived proteases of Betula verrucosa.
topic birch pollen
allergy
protease
proteome
transcriptome
zymogram
url https://www.mdpi.com/1422-0067/18/7/1433
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