Enzymatic Analysis of Yeast Cell Wall-Resident GAPDH and Its Secretion

Cohen, Michael J.; Philippe, Brianne; Lipke, Peter N.

doi:10.1128/msphere.01027-20

Cited by 5 publications

(4 citation statements)

References 63 publications

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“…The ability of enolase to be adsorbed to the surface of pathogenic cells was shown for Streptococcus pneumoniae [55] and C. albicans [11]. In S. cerevisiae yeast, the species closely related to N. glabratus, the secretion of GAPDH protein, which in a hydrolyzed form serves as an antibacterial peptide in the external environment, was demonstrated [24].…”

Section: Discussionmentioning

confidence: 99%

“…The occurrence of GAPDH on the cell surface and additional cell surface-dependent functions has been characterized, at least to some extent, for several bacterial pathogens [14][15][16][17][18][19][20][21], but only occasionally for pathogenic fungi. GAPDH was detected on C. albicans cells [22][23][24] and suggested to act as an adhesin, as deduced from the observed interactions with host proteins such as laminin and fibronectin [25]. GAPDH has also been proposed to contribute to the evasion of the host immune response, e.g., by binding to proteins that regulate the activity of the complement system [26].…”

Section: Introductionmentioning

confidence: 95%

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Glyceraldehyde 3-Phosphate Dehydrogenase on the Surface of Candida albicans and Nakaseomyces glabratus Cells—A Moonlighting Protein That Binds Human Vitronectin and Plasminogen and Can Adsorb to Pathogenic Fungal Cells via Major Adhesins Als3 and Epa6

Bednarek,

Satala,

Zawrotniak

et al. 2024

IJMS

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Candida albicans and other closely related pathogenic yeast-like fungi carry on their surface numerous loosely adsorbed “moonlighting proteins”—proteins that play evolutionarily conserved intracellular functions but also appear on the cell surface and exhibit additional functions, e.g., contributing to attachment to host tissues. In the current work, we characterized this “moonlighting” role for glyceraldehyde 3-phosphate dehydrogenase (GAPDH, EC 1.2.1.12) of C. albicans and Nakaseomyces glabratus. GAPDH was directly visualized on the cell surface of both species and shown to play a significant part in the total capacity of fungal cells to bind two selected human host proteins—vitronectin and plasminogen. Using purified proteins, both host proteins were found to tightly interact with GAPDH, with dissociation constants in an order of 10−8 M, as determined by bio-layer interferometry and surface plasmon resonance measurements. It was also shown that exogenous GAPDH tightly adheres to the surface of candidal cells, suggesting that the cell surface location of this moonlighting protein may partly result from the readsorption of its soluble form, which may be present at an infection site (e.g., due to release from dying fungal cells). The major dedicated adhesins, covalently bound to the cell wall—agglutinin-like sequence protein 3 (Als3) and epithelial adhesin 6 (Epa6)—were suggested to serve as the docking platforms for GAPDH in C. albicans and N. glabratus, respectively.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 95%

Glyceraldehyde 3-Phosphate Dehydrogenase on the Surface of Candida albicans and Nakaseomyces glabratus Cells—A Moonlighting Protein That Binds Human Vitronectin and Plasminogen and Can Adsorb to Pathogenic Fungal Cells via Major Adhesins Als3 and Epa6

Bednarek,

Satala,

Zawrotniak

et al. 2024

IJMS

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“…Beta-Glucan Quantification. The total cell wall extract was prepared as described by ref 84 with slight modifications. Briefly, the cells were incubated with Zymolyase at 37 °C for 1−2 h, followed by cell wall disruption by vortex.…”

Section: Measurement Of Surface Tension and Critical Micelle Concentr...mentioning

confidence: 99%

ASLdC3: A Derivative of Acidic Sophorolipid Disrupts Mitochondrial Function, Induces ROS Generation, and Inhibits Biofilm Formation in Candida albicans

Basotra,

Kumari,

Vij

et al. 2024

ACS Infect. Dis.

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Fungal infections account for more than 140 million cases of severe and life-threatening conditions each year, causing approximately 1.7 million deaths annually. Candida albicans and related species are the most common human fungal pathogens, causing both superficial (mucosal and cutaneous) and life-threatening invasive infections (candidemia) with a 40–75% mortality rate. Among many virulence factors of Candida albicans, morphological transition from yeast to hyphae, secretion of hydrolytic enzymes, and formation of biofilms are considered to be crucial for pathogenicity. However, the arsenals for the treatment against these pathogens are restricted to only a few classes of approved drugs, the efficacy of which is being compromised by host toxicity, fungistatic activity, and the emergence of drug resistance. In this study, we have described the development of a molecule, exhibiting excellent antifungal activity (MIC 8 μg/mL), by tailoring acidic sophorolipids with aryl alcohols via enzyme catalysis. This novel derivative, ASLdC3, is a surface-active compound that lowers the surface tension of the air–water interface up to 2-fold before reaching the critical micelle concentration of 25 μg/mL. ASLdC3 exhibits excellent antibiofilm properties against Candida albicans and other nonalbicans Candida species. The molecule primarily exhibits its antifungal activity by perturbing mitochondrial function through the alteration of the mitochondrial membrane potential (MMP) and generation of reactive oxygen species (ROS). The ROS damages fungal cell membrane function and cell wall integrity, eventually leading to cell death. ASLdC3 was found to be nontoxic in in vitro assay and nonhemolytic. Besides, it does not cause toxicity in the C. elegans model. Our study provides a valuable foundation for the potential of acidic sophorolipid as a nontoxic, biodegradable precursor for the design and synthesis of novel molecules for use as antimicrobial drugs as well as for other clinical applications.

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“…However, they are secreted outside the bacterial cell or attached to the cell membrane by an alternative mechanism and there they interact with host extracellular matrix (ECM) proteins [9]. Two recent works differentiate on cell wall proteomics-on whether yeast external GAPDH is a product of secretion or a cellular leakage, with nonconventional secretion being prevalent [10,11].…”

Section: Introductionmentioning

confidence: 99%

Pathogen Moonlighting Proteins: From Ancestral Key Metabolic Enzymes to Virulence Factors

et al. 2021

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Moonlighting and multitasking proteins refer to proteins with two or more functions performed by a single polypeptide chain. An amazing example of the Gain of Function (GoF) phenomenon of these proteins is that 25% of the moonlighting functions of our Multitasking Proteins Database (MultitaskProtDB-II) are related to pathogen virulence activity. Moreover, they usually have a canonical function belonging to highly conserved ancestral key functions, and their moonlighting functions are often involved in inducing extracellular matrix (ECM) protein remodeling. There are three main questions in the context of moonlighting proteins in pathogen virulence: (A) Why are a high percentage of pathogen moonlighting proteins involved in virulence? (B) Why do most of the canonical functions of these moonlighting proteins belong to primary metabolism? Moreover, why are they common in many pathogen species? (C) How are these different protein sequences and structures able to bind the same set of host ECM protein targets, mainly plasminogen (PLG), and colonize host tissues? By means of an extensive bioinformatics analysis, we suggest answers and approaches to these questions. There are three main ideas derived from the work: first, moonlighting proteins are not good candidates for vaccines. Second, several motifs that might be important in the adhesion to the ECM were identified. Third, an overrepresentation of GO codes related with virulence in moonlighting proteins were seen.

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Enzymatic Analysis of Yeast Cell Wall-Resident GAPDH and Its Secretion

Cited by 5 publications

References 63 publications

Glyceraldehyde 3-Phosphate Dehydrogenase on the Surface of Candida albicans and Nakaseomyces glabratus Cells—A Moonlighting Protein That Binds Human Vitronectin and Plasminogen and Can Adsorb to Pathogenic Fungal Cells via Major Adhesins Als3 and Epa6

Glyceraldehyde 3-Phosphate Dehydrogenase on the Surface of Candida albicans and Nakaseomyces glabratus Cells—A Moonlighting Protein That Binds Human Vitronectin and Plasminogen and Can Adsorb to Pathogenic Fungal Cells via Major Adhesins Als3 and Epa6

ASLdC3: A Derivative of Acidic Sophorolipid Disrupts Mitochondrial Function, Induces ROS Generation, and Inhibits Biofilm Formation in Candida albicans

Pathogen Moonlighting Proteins: From Ancestral Key Metabolic Enzymes to Virulence Factors

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