Heterologous expression and biochemical characterization of an α1,2-mannosidase encoded by the Candida albicans MNS1 gene
Protein glycosylation pathways, commonly found in fungal pathogens, offer an attractive new area of study for the discovery of antifungal targets. In particular, these post-translational modifications are required for virulence and proper cell wall assembly in Candida albicans, an opportunistic huma...
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2008
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ftdoajarticles:oai:doaj.org/article:68d91473ca814bb399f557e0e6ae32f3 2023-05-15T15:09:31+02:00 Heterologous expression and biochemical characterization of an α1,2-mannosidase encoded by the Candida albicans MNS1 gene Héctor M Mora-Montes Everardo López-Romero Samuel Zinker Patricia Ponce-Noyola Arturo Flores-Carreón 2008-11-01T00:00:00Z https://doi.org/10.1590/S0074-02762008000700016 https://doaj.org/article/68d91473ca814bb399f557e0e6ae32f3 EN eng Instituto Oswaldo Cruz, Ministério da Saúde http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762008000700016 https://doaj.org/toc/0074-0276 https://doaj.org/toc/1678-8060 doi:10.1590/S0074-02762008000700016 0074-0276 1678-8060 https://doaj.org/article/68d91473ca814bb399f557e0e6ae32f3 Memórias do Instituto Oswaldo Cruz., Vol 103, Iss 7, Pp 724-730 (2008) 1,2-mannosidase heterologous expression glycosyl hydrolase family 47 Candida albicans polyclonal antibodies N-glycosylation Arctic medicine. Tropical medicine RC955-962 Microbiology QR1-502 article 2008 ftdoajarticles https://doi.org/10.1590/S0074-02762008000700016 2023-01-08T01:33:41Z Protein glycosylation pathways, commonly found in fungal pathogens, offer an attractive new area of study for the discovery of antifungal targets. In particular, these post-translational modifications are required for virulence and proper cell wall assembly in Candida albicans, an opportunistic human pathogen. The C. albicans MNS1 gene is predicted to encode a member of the glycosyl hydrolase family 47, with 1,2-mannosidase activity. In order to characterise its activity, we first cloned the C. albicans MNS1 gene into Escherichia coli, then expressed and purified the enzyme. The recombinant Mns1 was capable of converting a Man9GlcNAc2 N-glycan core into Man8GlcNAc2 isomer B, but failed to process a Man5GlcNAc2-Asn N-oligosaccharide. These properties are similar to those displayed by Mns1 purified from C. albicansmembranes and strongly suggest that the enzyme is an ±1,2-mannosidase that is localised to the endoplasmic reticulum and involved in the processing of N-linked mannans. Polyclonal antibodies specifically raised against recombinant Mns1 also immunoreacted with the soluble ±1,2-mannosidases E-I and E-II, indicating that Mns1 could share structural similarities with both soluble enzymes. Due to the high degree of similarity between the members of family 47, it is conceivable that these antibodies may recognise ±1,2-mannosidases in other biological systems as well. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Memórias do Instituto Oswaldo Cruz 103 7 724 730 |
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1,2-mannosidase heterologous expression glycosyl hydrolase family 47 Candida albicans polyclonal antibodies N-glycosylation Arctic medicine. Tropical medicine RC955-962 Microbiology QR1-502 |
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1,2-mannosidase heterologous expression glycosyl hydrolase family 47 Candida albicans polyclonal antibodies N-glycosylation Arctic medicine. Tropical medicine RC955-962 Microbiology QR1-502 Héctor M Mora-Montes Everardo López-Romero Samuel Zinker Patricia Ponce-Noyola Arturo Flores-Carreón Heterologous expression and biochemical characterization of an α1,2-mannosidase encoded by the Candida albicans MNS1 gene |
topic_facet |
1,2-mannosidase heterologous expression glycosyl hydrolase family 47 Candida albicans polyclonal antibodies N-glycosylation Arctic medicine. Tropical medicine RC955-962 Microbiology QR1-502 |
description |
Protein glycosylation pathways, commonly found in fungal pathogens, offer an attractive new area of study for the discovery of antifungal targets. In particular, these post-translational modifications are required for virulence and proper cell wall assembly in Candida albicans, an opportunistic human pathogen. The C. albicans MNS1 gene is predicted to encode a member of the glycosyl hydrolase family 47, with 1,2-mannosidase activity. In order to characterise its activity, we first cloned the C. albicans MNS1 gene into Escherichia coli, then expressed and purified the enzyme. The recombinant Mns1 was capable of converting a Man9GlcNAc2 N-glycan core into Man8GlcNAc2 isomer B, but failed to process a Man5GlcNAc2-Asn N-oligosaccharide. These properties are similar to those displayed by Mns1 purified from C. albicansmembranes and strongly suggest that the enzyme is an ±1,2-mannosidase that is localised to the endoplasmic reticulum and involved in the processing of N-linked mannans. Polyclonal antibodies specifically raised against recombinant Mns1 also immunoreacted with the soluble ±1,2-mannosidases E-I and E-II, indicating that Mns1 could share structural similarities with both soluble enzymes. Due to the high degree of similarity between the members of family 47, it is conceivable that these antibodies may recognise ±1,2-mannosidases in other biological systems as well. |
format |
Article in Journal/Newspaper |
author |
Héctor M Mora-Montes Everardo López-Romero Samuel Zinker Patricia Ponce-Noyola Arturo Flores-Carreón |
author_facet |
Héctor M Mora-Montes Everardo López-Romero Samuel Zinker Patricia Ponce-Noyola Arturo Flores-Carreón |
author_sort |
Héctor M Mora-Montes |
title |
Heterologous expression and biochemical characterization of an α1,2-mannosidase encoded by the Candida albicans MNS1 gene |
title_short |
Heterologous expression and biochemical characterization of an α1,2-mannosidase encoded by the Candida albicans MNS1 gene |
title_full |
Heterologous expression and biochemical characterization of an α1,2-mannosidase encoded by the Candida albicans MNS1 gene |
title_fullStr |
Heterologous expression and biochemical characterization of an α1,2-mannosidase encoded by the Candida albicans MNS1 gene |
title_full_unstemmed |
Heterologous expression and biochemical characterization of an α1,2-mannosidase encoded by the Candida albicans MNS1 gene |
title_sort |
heterologous expression and biochemical characterization of an α1,2-mannosidase encoded by the candida albicans mns1 gene |
publisher |
Instituto Oswaldo Cruz, Ministério da Saúde |
publishDate |
2008 |
url |
https://doi.org/10.1590/S0074-02762008000700016 https://doaj.org/article/68d91473ca814bb399f557e0e6ae32f3 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Memórias do Instituto Oswaldo Cruz., Vol 103, Iss 7, Pp 724-730 (2008) |
op_relation |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762008000700016 https://doaj.org/toc/0074-0276 https://doaj.org/toc/1678-8060 doi:10.1590/S0074-02762008000700016 0074-0276 1678-8060 https://doaj.org/article/68d91473ca814bb399f557e0e6ae32f3 |
op_doi |
https://doi.org/10.1590/S0074-02762008000700016 |
container_title |
Memórias do Instituto Oswaldo Cruz |
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103 |
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7 |
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724 |
op_container_end_page |
730 |
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1766340692641054720 |