The Heat Shock Protein 60 and Pap1 Participate in the Sporothrix schenckii-Host Interaction

Sporothrixschenckii is one of the etiological agents of sporotrichosis, a worldwide-distributed subcutaneous mycosis. Its cell wall contains a glycoconjugate composed of rhamnose, mannose, glucuronic acid, and proteins, named peptidorhamnomannan, which harbors i...

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Autores principales:	Laura C. García-Carnero, Roberta Salinas-Marín, Nancy E. Lozoya-Pérez, Katarzyna Wrobel, Kazimierz Wrobel, Iván Martínez-Duncker, Gustavo A. Niño-Vega, Héctor M. Mora-Montes
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	sporotrichosis cell wall adhesin glycoprotein recombinant protein virulence Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/d8fb30fe6185445fb226b74c2e46c313
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spelling	oai:doaj.org-article:d8fb30fe6185445fb226b74c2e46c3132021-11-25T18:06:12ZThe Heat Shock Protein 60 and Pap1 Participate in the <i>Sporothrix</i> <i>schenckii</i>-Host Interaction10.3390/jof71109602309-608Xhttps://doaj.org/article/d8fb30fe6185445fb226b74c2e46c3132021-11-01T00:00:00Zhttps://www.mdpi.com/2309-608X/7/11/960https://doaj.org/toc/2309-608X<i>Sporothrix</i><i>schenckii</i> is one of the etiological agents of sporotrichosis, a worldwide-distributed subcutaneous mycosis. Its cell wall contains a glycoconjugate composed of rhamnose, mannose, glucuronic acid, and proteins, named peptidorhamnomannan, which harbors important <i>Sporothrix</i>-specific immunogenic epitopes. Although the peptidorhamnomannan carbohydrate moiety has been extensively studied, thus far, little is known about the protein core. Here, using LC-MS/MS, we analyzed the <i>S.</i><i>schenckii</i> peptidorhamnomannan peptide fraction and generated mass signals of 325 proteins, most of them likely to be moonlighting proteins. Among the identified proteins, chaperonin GroEL/Hsp60 and the uncharacterized protein Pap1 were selected for further analysis. Both proteins were heterologously expressed in bacteria, and they showed adhesive properties to the extracellular matrix proteins laminin, elastin, fibrinogen, and fibronectin, although Pap1 also was bound to type-I and type-II collagen. The inoculation of concentrations higher than 40 μg of these proteins, separately, increased immune effectors in the hemolymph of <i>Galleria</i><i>mellonella</i> larvae and protected animals from an <i>S.</i><i>schenckii</i> lethal challenge. These observations were confirmed when yeast-like cells, pre-incubated with anti-rHsp60 or anti-rPap1 antibodies were used to inoculate larvae. The animals inoculated with pretreated cells showed increased survival rates when compared to the control groups. In conclusion, we report that Hsp60 and Pap1 are part of the cell wall peptidorhamnomannan, can bind extracellular matrix components, and contribute to the <i>S.</i><i>schenckii</i> virulence. To our knowledge, this is the first report about moonlighting protein in the <i>S.</i><i>schenckii</i> cell wall with an important role during the pathogen–host interaction.Laura C. García-CarneroRoberta Salinas-MarínNancy E. Lozoya-PérezKatarzyna WrobelKazimierz WrobelIván Martínez-DunckerGustavo A. Niño-VegaHéctor M. Mora-MontesMDPI AGarticlesporotrichosiscell walladhesinglycoproteinrecombinant proteinvirulenceBiology (General)QH301-705.5ENJournal of Fungi, Vol 7, Iss 960, p 960 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	sporotrichosis cell wall adhesin glycoprotein recombinant protein virulence Biology (General) QH301-705.5
spellingShingle	sporotrichosis cell wall adhesin glycoprotein recombinant protein virulence Biology (General) QH301-705.5 Laura C. García-Carnero Roberta Salinas-Marín Nancy E. Lozoya-Pérez Katarzyna Wrobel Kazimierz Wrobel Iván Martínez-Duncker Gustavo A. Niño-Vega Héctor M. Mora-Montes The Heat Shock Protein 60 and Pap1 Participate in the <i>Sporothrix</i> <i>schenckii</i>-Host Interaction
description	<i>Sporothrix</i><i>schenckii</i> is one of the etiological agents of sporotrichosis, a worldwide-distributed subcutaneous mycosis. Its cell wall contains a glycoconjugate composed of rhamnose, mannose, glucuronic acid, and proteins, named peptidorhamnomannan, which harbors important <i>Sporothrix</i>-specific immunogenic epitopes. Although the peptidorhamnomannan carbohydrate moiety has been extensively studied, thus far, little is known about the protein core. Here, using LC-MS/MS, we analyzed the <i>S.</i><i>schenckii</i> peptidorhamnomannan peptide fraction and generated mass signals of 325 proteins, most of them likely to be moonlighting proteins. Among the identified proteins, chaperonin GroEL/Hsp60 and the uncharacterized protein Pap1 were selected for further analysis. Both proteins were heterologously expressed in bacteria, and they showed adhesive properties to the extracellular matrix proteins laminin, elastin, fibrinogen, and fibronectin, although Pap1 also was bound to type-I and type-II collagen. The inoculation of concentrations higher than 40 μg of these proteins, separately, increased immune effectors in the hemolymph of <i>Galleria</i><i>mellonella</i> larvae and protected animals from an <i>S.</i><i>schenckii</i> lethal challenge. These observations were confirmed when yeast-like cells, pre-incubated with anti-rHsp60 or anti-rPap1 antibodies were used to inoculate larvae. The animals inoculated with pretreated cells showed increased survival rates when compared to the control groups. In conclusion, we report that Hsp60 and Pap1 are part of the cell wall peptidorhamnomannan, can bind extracellular matrix components, and contribute to the <i>S.</i><i>schenckii</i> virulence. To our knowledge, this is the first report about moonlighting protein in the <i>S.</i><i>schenckii</i> cell wall with an important role during the pathogen–host interaction.
format	article
author	Laura C. García-Carnero Roberta Salinas-Marín Nancy E. Lozoya-Pérez Katarzyna Wrobel Kazimierz Wrobel Iván Martínez-Duncker Gustavo A. Niño-Vega Héctor M. Mora-Montes
author_facet	Laura C. García-Carnero Roberta Salinas-Marín Nancy E. Lozoya-Pérez Katarzyna Wrobel Kazimierz Wrobel Iván Martínez-Duncker Gustavo A. Niño-Vega Héctor M. Mora-Montes
author_sort	Laura C. García-Carnero
title	The Heat Shock Protein 60 and Pap1 Participate in the <i>Sporothrix</i> <i>schenckii</i>-Host Interaction
title_short	The Heat Shock Protein 60 and Pap1 Participate in the <i>Sporothrix</i> <i>schenckii</i>-Host Interaction
title_full	The Heat Shock Protein 60 and Pap1 Participate in the <i>Sporothrix</i> <i>schenckii</i>-Host Interaction
title_fullStr	The Heat Shock Protein 60 and Pap1 Participate in the <i>Sporothrix</i> <i>schenckii</i>-Host Interaction
title_full_unstemmed	The Heat Shock Protein 60 and Pap1 Participate in the <i>Sporothrix</i> <i>schenckii</i>-Host Interaction
title_sort	heat shock protein 60 and pap1 participate in the <i>sporothrix</i> <i>schenckii</i>-host interaction
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/d8fb30fe6185445fb226b74c2e46c313
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The Heat Shock Protein 60 and Pap1 Participate in the <i>Sporothrix</i> <i>schenckii</i>-Host Interaction

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