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Cell surface binding strategy of Bgl2/Bgl1-orthgroup proteins differs from strategies deployed by other fungal glucanases of GH17-familyтезисы доклада
- Авторы: Sabirzianov F., Rekstina V., Sabirzianova T., Kudriashova I., Kalebina T.
- Сборник: Abstract Book, Fungal Cell Wall 2015 Conference
- Тезисы
- Год издания: 2015
- Место издания: Institut Pasteur Paris, France
- Первая страница: 68
- Последняя страница: 68
- Аннотация: Objective: Ascomycete pathogen's cell wall (CW) mainly consists of glucan, which is synthesized as a linear polymer and then is modified by branching enzymes localized on the cell surface. According to domain architecture most of fungal GH17-family branching glucanases use strong cell surface anchoring strategies via TM-domain, GPI-anchor or alkali sensitive linkage. Highly conserved Bgl2/Bgt1-orthogroup GH17-family glucanases do not have such domains. It is not clear what binding strategy is used by Bgl2/Bgt1-orthogroup. This knowledge is important for investigation of pathogen-host interactions, becauseBgl2 is highly immunogenic and may serve as diagnostic and prognostic marker of infections causedby Candida albicans (Pitarch et al, 2006). Methods: Bioinformatic analysis, PCR-mutagenesis, Western blotting, electron microscopy and non-pathogenic Saccharomyces cerevisiae strain BY4742 as a model were used. Results: Basing on comparison of fungal GH17-family proteins we have predicted that C-terminal region of Bgl2 may participate in the attachment of Bgl2 to the CW. Comparative analysis after deletion of this region demonstrated, that wild type Bgl2 was present in CW after 1% SDS and trypsin treatment and could be effectively extracted only in hot Laemmli sample buffer or 6M guanidine chloride, while Bgl2 lacking C-terminal region was removed by SDS treatment. Fibrils were observed in Bgl2 extracts from wild type CW, but not in extracts from mutant CW. Conclusions: We have demonstrated that Bgl2 was extremely strongly anchored to the CW via formation of structures similar to amyloid like fibrils. C-terminal region of Bgl2was involved it in the attachment to the CW.
- Добавил в систему: Кудряшова Ирина Борисовна