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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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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