Global Identification of Biofilm-Specific Proteolysis in <named-content content-type="genus-species">Candida albicans</named-content>

ABSTRACT Candida albicans is a fungal species that is part of the normal human microbiota and also an opportunistic pathogen capable of causing mucosal and systemic infections. C. albicans cells proliferate in a planktonic (suspension) state, but they also form biofilms, organized and tightly packed...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Michael B. Winter, Eugenia C. Salcedo, Matthew B. Lohse, Nairi Hartooni, Megha Gulati, Hiram Sanchez, Julie Takagi, Bernhard Hube, David R. Andes, Alexander D. Johnson, Charles S. Craik, Clarissa J. Nobile
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2016
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/b251c013752b4e168b6bd0d6f5a3ae7d
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:b251c013752b4e168b6bd0d6f5a3ae7d
record_format	dspace
spelling	oai:doaj.org-article:b251c013752b4e168b6bd0d6f5a3ae7d2021-11-15T15:50:15ZGlobal Identification of Biofilm-Specific Proteolysis in <named-content content-type="genus-species">Candida albicans</named-content>10.1128/mBio.01514-162150-7511https://doaj.org/article/b251c013752b4e168b6bd0d6f5a3ae7d2016-11-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01514-16https://doaj.org/toc/2150-7511ABSTRACT Candida albicans is a fungal species that is part of the normal human microbiota and also an opportunistic pathogen capable of causing mucosal and systemic infections. C. albicans cells proliferate in a planktonic (suspension) state, but they also form biofilms, organized and tightly packed communities of cells attached to a solid surface. Biofilms colonize many niches of the human body and persist on implanted medical devices, where they are a major source of new C. albicans infections. Here, we used an unbiased and global substrate-profiling approach to discover proteolytic activities produced specifically by C. albicans biofilms, compared to planktonic cells, with the goal of identifying potential biofilm-specific diagnostic markers and targets for therapeutic intervention. This activity-based profiling approach, coupled with proteomics, identified Sap5 (Candidapepsin-5) and Sap6 (Candidapepsin-6) as major biofilm-specific proteases secreted by C. albicans. Fluorogenic peptide substrates with selectivity for Sap5 or Sap6 confirmed that their activities are highly upregulated in C. albicans biofilms; we also show that these activities are upregulated in other Candida clade pathogens. Deletion of the SAP5 and SAP6 genes in C. albicans compromised biofilm development in vitro in standard biofilm assays and in vivo in a rat central venous catheter biofilm model. This work establishes secreted proteolysis as a promising enzymatic marker and potential therapeutic target for Candida biofilm formation. IMPORTANCE Biofilm formation by the opportunistic fungal pathogen C. albicans is a major cause of life-threatening infections. This work provides a global characterization of secreted proteolytic activity produced specifically by C. albicans biofilms. We identify activity from the proteases Sap5 and Sap6 as highly upregulated during C. albicans biofilm formation and develop Sap-cleavable fluorogenic substrates that enable the detection of biofilms from C. albicans and also from additional pathogenic Candida species. Furthermore, SAP5 and SAP6 deletions confirm that both proteases are required for proper biofilm development in vitro and in vivo. We propose that secreted proteolysis is a promising marker for the diagnosis and potential therapeutic targeting of Candida biofilm-associated infections.Michael B. WinterEugenia C. SalcedoMatthew B. LohseNairi HartooniMegha GulatiHiram SanchezJulie TakagiBernhard HubeDavid R. AndesAlexander D. JohnsonCharles S. CraikClarissa J. NobileAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 7, Iss 5 (2016)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Michael B. Winter Eugenia C. Salcedo Matthew B. Lohse Nairi Hartooni Megha Gulati Hiram Sanchez Julie Takagi Bernhard Hube David R. Andes Alexander D. Johnson Charles S. Craik Clarissa J. Nobile Global Identification of Biofilm-Specific Proteolysis in <named-content content-type="genus-species">Candida albicans</named-content>
description	ABSTRACT Candida albicans is a fungal species that is part of the normal human microbiota and also an opportunistic pathogen capable of causing mucosal and systemic infections. C. albicans cells proliferate in a planktonic (suspension) state, but they also form biofilms, organized and tightly packed communities of cells attached to a solid surface. Biofilms colonize many niches of the human body and persist on implanted medical devices, where they are a major source of new C. albicans infections. Here, we used an unbiased and global substrate-profiling approach to discover proteolytic activities produced specifically by C. albicans biofilms, compared to planktonic cells, with the goal of identifying potential biofilm-specific diagnostic markers and targets for therapeutic intervention. This activity-based profiling approach, coupled with proteomics, identified Sap5 (Candidapepsin-5) and Sap6 (Candidapepsin-6) as major biofilm-specific proteases secreted by C. albicans. Fluorogenic peptide substrates with selectivity for Sap5 or Sap6 confirmed that their activities are highly upregulated in C. albicans biofilms; we also show that these activities are upregulated in other Candida clade pathogens. Deletion of the SAP5 and SAP6 genes in C. albicans compromised biofilm development in vitro in standard biofilm assays and in vivo in a rat central venous catheter biofilm model. This work establishes secreted proteolysis as a promising enzymatic marker and potential therapeutic target for Candida biofilm formation. IMPORTANCE Biofilm formation by the opportunistic fungal pathogen C. albicans is a major cause of life-threatening infections. This work provides a global characterization of secreted proteolytic activity produced specifically by C. albicans biofilms. We identify activity from the proteases Sap5 and Sap6 as highly upregulated during C. albicans biofilm formation and develop Sap-cleavable fluorogenic substrates that enable the detection of biofilms from C. albicans and also from additional pathogenic Candida species. Furthermore, SAP5 and SAP6 deletions confirm that both proteases are required for proper biofilm development in vitro and in vivo. We propose that secreted proteolysis is a promising marker for the diagnosis and potential therapeutic targeting of Candida biofilm-associated infections.
format	article
author	Michael B. Winter Eugenia C. Salcedo Matthew B. Lohse Nairi Hartooni Megha Gulati Hiram Sanchez Julie Takagi Bernhard Hube David R. Andes Alexander D. Johnson Charles S. Craik Clarissa J. Nobile
author_facet	Michael B. Winter Eugenia C. Salcedo Matthew B. Lohse Nairi Hartooni Megha Gulati Hiram Sanchez Julie Takagi Bernhard Hube David R. Andes Alexander D. Johnson Charles S. Craik Clarissa J. Nobile
author_sort	Michael B. Winter
title	Global Identification of Biofilm-Specific Proteolysis in <named-content content-type="genus-species">Candida albicans</named-content>
title_short	Global Identification of Biofilm-Specific Proteolysis in <named-content content-type="genus-species">Candida albicans</named-content>
title_full	Global Identification of Biofilm-Specific Proteolysis in <named-content content-type="genus-species">Candida albicans</named-content>
title_fullStr	Global Identification of Biofilm-Specific Proteolysis in <named-content content-type="genus-species">Candida albicans</named-content>
title_full_unstemmed	Global Identification of Biofilm-Specific Proteolysis in <named-content content-type="genus-species">Candida albicans</named-content>
title_sort	global identification of biofilm-specific proteolysis in <named-content content-type="genus-species">candida albicans</named-content>
publisher	American Society for Microbiology
publishDate	2016
url	https://doaj.org/article/b251c013752b4e168b6bd0d6f5a3ae7d
work_keys_str_mv	AT michaelbwinter globalidentificationofbiofilmspecificproteolysisinnamedcontentcontenttypegenusspeciescandidaalbicansnamedcontent AT eugeniacsalcedo globalidentificationofbiofilmspecificproteolysisinnamedcontentcontenttypegenusspeciescandidaalbicansnamedcontent AT matthewblohse globalidentificationofbiofilmspecificproteolysisinnamedcontentcontenttypegenusspeciescandidaalbicansnamedcontent AT nairihartooni globalidentificationofbiofilmspecificproteolysisinnamedcontentcontenttypegenusspeciescandidaalbicansnamedcontent AT meghagulati globalidentificationofbiofilmspecificproteolysisinnamedcontentcontenttypegenusspeciescandidaalbicansnamedcontent AT hiramsanchez globalidentificationofbiofilmspecificproteolysisinnamedcontentcontenttypegenusspeciescandidaalbicansnamedcontent AT julietakagi globalidentificationofbiofilmspecificproteolysisinnamedcontentcontenttypegenusspeciescandidaalbicansnamedcontent AT bernhardhube globalidentificationofbiofilmspecificproteolysisinnamedcontentcontenttypegenusspeciescandidaalbicansnamedcontent AT davidrandes globalidentificationofbiofilmspecificproteolysisinnamedcontentcontenttypegenusspeciescandidaalbicansnamedcontent AT alexanderdjohnson globalidentificationofbiofilmspecificproteolysisinnamedcontentcontenttypegenusspeciescandidaalbicansnamedcontent AT charlesscraik globalidentificationofbiofilmspecificproteolysisinnamedcontentcontenttypegenusspeciescandidaalbicansnamedcontent AT clarissajnobile globalidentificationofbiofilmspecificproteolysisinnamedcontentcontenttypegenusspeciescandidaalbicansnamedcontent
_version_	1718427479247421440

Global Identification of Biofilm-Specific Proteolysis in <named-content content-type="genus-species">Candida albicans</named-content>

Ejemplares similares