Isolation of Sporothrix schenckii MNT1 and the biochemical and functional characterization of the encoded α1,2-mannosyltransferase activity

Hernández‐Cervantes, Arturo; Mora-Montes, Héctor M.; Álvarez-Vargas, Aurelio; Jiménez, Diana F. Díaz; Robledo-Ortiz, Claudia I; Flores-Carreón, Arturo

doi:10.1099/mic.0.060392-0

Cited by 15 publications

(5 citation statements)

References 51 publications

(43 reference statements)

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“…In agreement, the interaction of H. capsulatum , P. brasiliensis , C. neoformans , C. albicans , and S. cerevisiae with Ac could also be inhibited by soluble mannose, correlating with the presence of high amounts of mannans on the surface of these fungi, whereas in the case of the fungus S. brasiliensis , either absence of free‐mannan on its surface or participation of other interaction receptors cannot be ruled out (Hernandez‐Cervantes et al, ; Lopes‐Bezerra et al, ).…”

Section: Discussionmentioning

confidence: 78%

Unravelling the interactions of the environmental hostAcanthamoeba castellaniiwith fungi through the recognition by mannose‐binding proteins

Gonçalves

Ferreira

Gomes

et al. 2019

Cellular Microbiology

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Free-living amoebae (FLAs) are major reservoirs for a variety of bacteria, viruses, and fungi. The most studied mycophagic FLA, Acanthamoeba castellanii (Ac), is a potential environmental host for endemic fungal pathogens such as Cryptococcus spp., Histoplasma capsulatum, Blastomyces dermatitides, and Sporothrix schenckii. However, the mechanisms involved in this interaction are poorly understood. The aim of this

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Section: Discussionmentioning

confidence: 78%

Unravelling the interactions of the environmental hostAcanthamoeba castellaniiwith fungi through the recognition by mannose‐binding proteins

Gonçalves

Ferreira

Gomes

et al. 2019

Cellular Microbiology

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“…The samples were loaded onto a 4% PAGE gel and run for 11 h at 40 V under native conditions. The N -acetylhexosaminidase activity was determined by incubating with 0.4 mM 4-methylumbelliferyl N-acetyl-β-D-glucosamine (Sigma) in 0.1 M citrate-KOH buffer (pH 4.5) during 30 min at 37°C, and the results were observed by exposing the gel to UV light (Hernandez-Cervantes et al, 2012 ).…”

Section: Methodsmentioning

confidence: 99%

Role of Protein Glycosylation in Candida parapsilosis Cell Wall Integrity and Host Interaction

et al. 2016

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Candida parapsilosis is an important, emerging opportunistic fungal pathogen. Highly mannosylated fungal cell wall proteins are initial contact points with host immune systems. In Candida albicans, Och1 is a Golgi α1,6-mannosyltransferase that plays a key role in the elaboration of the N-linked mannan outer chain. Here, we disrupted C. parapsilosis OCH1 to gain insights into the contribution of N-linked mannosylation to cell fitness and to interactions with immune cells. Loss of Och1 in C. parapsilosis resulted in cellular aggregation, failure of morphogenesis, enhanced susceptibility to cell wall perturbing agents and defects in wall composition. We removed the cell wall O-linked mannans by β-elimination, and assessed the relevance of mannans during interaction with human monocytes. Results indicated that O-linked mannans are important for IL-1β stimulation in a dectin-1 and TLR4-dependent pathway; whereas both, N- and O-linked mannans are equally important ligands for TNFα and IL-6 stimulation, but neither is involved in IL-10 production. Furthermore, mice infected with C. parapsilosis och1Δ null mutant cells had significantly lower fungal burdens compared to wild-type (WT)-challenged counterparts. Therefore, our data are the first to demonstrate that C. parapsilosis N- and O-linked mannans have different roles in host interactions than those reported for C. albicans.

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“…Samples containing 100 µg of total protein were loaded onto a 6% PAGE gel and run for 3 h at 70 V under native conditions. The β-N-acetylhexosaminidase activity was determined by incubating with 0.4 mM 4-methylumbelliferyl N-acetyl-β-d-glucosamine (Sigma) in 0.1 M citrate-KOH buffer, pH 4.5 for 30 min at 37 • C. The enzyme activity was revealed by exposing the gels to UV light [43]. In some experiments, the cell homogenates were N-linked deglycosylated by incubating the samples for 20 h at 37 • C with 25 U of endoglycosidase H, prior to the electrophoretic separation [31].…”

Section: Electrophoretic Mobility Shift Assays Of Hex1mentioning

confidence: 99%

“…The C. albicans Hex1, a β-N-acetylhexosaminidase, is a highly N-linked mannosylated protein that has been previously used to show defects in the N-linked mannosylation pathway [29,31,49]. Therefore, we performed non-denaturing protein electrophoresis followed by an in situ zymogram with a fluorogenic substrate [43]. The protein from the kex2∆ null mutant, but not that from the WT or the reintegrant control strains, had increased electrophoretic mobility, which suggested the presence of shorter N-linked mannans ( Figure 2B).…”

Section: Loss Of Kex2 Affects the C Albicans Cell Wall And Protein Gmentioning

confidence: 99%

Loss of Kex2 Affects the Candida albicans Cell Wall and Interaction with Innate Immune Cells

Gómez-Gaviria

Lozoya-Pérez

Staniszewska

et al. 2020

JoF

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The secretory pathway in Candida albicans involves the protein translocation into the lumen of the endoplasmic reticulum and transport to the Golgi complex, where proteins undergo posttranslational modifications, including glycosylation and proteolysis. The Golgi-resident Kex2 protease is involved in such processing and disruption of its encoding gene affected virulence and dimorphism. These previous studies were performed using cells without URA3 or with URA3 ectopically placed into the KEX2 locus. Since these conditions are known to affect the cellular fitness and the host–fungus interaction, here we generated a kex2Δ null mutant strain with URA3 placed into the neutral locus RPS1. The characterization of this strain showed defects in the cell wall composition, with a reduction in the N-linked mannan content, and the increment in the levels of O-linked mannans, chitin, and β-glucans. The defects in the mannan content are likely linked to changes in Golgi-resident enzymes, as the α-1,2-mannosyltransferase and α-1,6-mannosyltransferase activities were incremented and reduced, respectively. The mutant cells also showed reduced ability to stimulate cytokine production and phagocytosis by human mononuclear cells and macrophages, respectively. Collectively, these data showed that loss of Kex2 affected the cell wall composition, the protein glycosylation pathways, and interaction with innate immune cells.

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Isolation of Sporothrix schenckii MNT1 and the biochemical and functional characterization of the encoded α1,2-mannosyltransferase activity

Cited by 15 publications

References 51 publications

Unravelling the interactions of the environmental hostAcanthamoeba castellaniiwith fungi through the recognition by mannose‐binding proteins

Unravelling the interactions of the environmental hostAcanthamoeba castellaniiwith fungi through the recognition by mannose‐binding proteins

Role of Protein Glycosylation in Candida parapsilosis Cell Wall Integrity and Host Interaction

Loss of Kex2 Affects the Candida albicans Cell Wall and Interaction with Innate Immune Cells

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