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Bacteriophage-encoded virion-associated enzymes to overcome the carbohydrate barriers during the infection process.

ABSTRACT: Bacteriophages are bacterial viruses that infect the host after successful receptor recognition and adsorption to the cell surface. The irreversible adherence followed by genome material ejection into host cell cytoplasm must be preceded by the passage of diverse carbohydrate barriers such as capsule polysaccharides (CPSs), O-polysaccharide chains of lipopolysaccharide (LPS) molecules, extracellular polysaccharides (EPSs) forming biofilm matrix, and peptidoglycan (PG) layers. For that purpose, bacteriophages are equipped with various virion-associated carbohydrate active enzymes, termed polysaccharide depolymerases and lysins, that recognize, bind, and degrade the polysaccharide compounds. We discuss the existing diversity in structural locations, variable architectures, enzymatic specificities, and evolutionary aspects of polysaccharide depolymerases and virion-associated lysins (VALs) and illustrate how these aspects can correlate with the host spectrum. In addition, we present methods that can be used for activity determination and the application potential of these enzymes as antibacterials, antivirulence agents, and diagnostic tools.

SUBMITTER: Latka A

PROVIDER: S-EPMC5380687 | biostudies-literature | 2017 Apr

REPOSITORIES: biostudies-literature

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Bacteriophage-encoded virion-associated enzymes to overcome the carbohydrate barriers during the infection process.

Latka Agnieszka A Maciejewska Barbara B Majkowska-Skrobek Grazyna G Briers Yves Y Drulis-Kawa Zuzanna Z

Applied microbiology and biotechnology 20170323 8

Bacteriophages are bacterial viruses that infect the host after successful receptor recognition and adsorption to the cell surface. The irreversible adherence followed by genome material ejection into host cell cytoplasm must be preceded by the passage of diverse carbohydrate barriers such as capsule polysaccharides (CPSs), O-polysaccharide chains of lipopolysaccharide (LPS) molecules, extracellular polysaccharides (EPSs) forming biofilm matrix, and peptidoglycan (PG) layers. For that purpose, b ...[more]

PMID: 28337580

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Project description:BackgroundIn emergencies, language barriers may have dangerous consequences for the patients. There have been some technical approaches to overcome language barriers in medical care but not yet in the prehospital emergency care setting. The use of digital technologies in health care is expanding rapidly. Involving end users at all stages of the development process may help to ensure such technologies are usable and can be implemented.ObjectiveWe aimed to develop a digital communication tool that addresses paramedic needs in the specific circumstances of prehospital emergency care and helps paramedics to overcome language barriers when providing care to foreign-language patients.MethodsWe actively engaged paramedics and software designers in an action-oriented, participatory, iterative development process, which included field observations, workshops, background conversations, questionnaires on rescue missions, studying the literature, and preliminary testing in the field.ResultsWith input from paramedics, we created an app with 600 fixed phrases supporting 18 languages. The app includes medical history-taking questions, phrases asking for consent, and phrases providing specific additional information. Children as patients, as well as their carers and other third parties, can be addressed with appropriate wording. All phrases can be played back audibly or displayed as text. The comprehensive content is grouped into categories and adapted to diverse scenarios, which makes the tool rapidly usable. The app includes a function to document patient responses and the conversation history. For evaluation in a clinical study, the app is run on a smartphone with extra speakers to be of use in noisy environments. The use of prototypes proved valuable to verify that the content, structure, and functions discussed in theory were of value and genuinely needed in practice and that the various device control elements were intuitive.ConclusionsThe nature of the paramedic work environment places specific demands on the communication options used and need for such devices. The active involvement of paramedics in the development process allowed us to understand and subsequently consider their experience-based knowledge. Software designers could understand the paramedics' work environment and consider respective needs in the menu navigation and design principles of the app. We argue that the development of any medical software product should actively involve both end users and developers in all phases of the development process. Providing the users with the opportunity to influence technology development ensures that the result is closer to their needs, which can be seen as crucial for successful implementation and sustainable use.Trial registrationGerman Clinical Trials Register DRKS00016719; https://www.drks.de/drks_web/navigate.do?navigationId=trial.HTML&TRIAL_ID=DRKS00016719.International registered report identifier (irrid)RR2-10.1186/s12913-020-05098-5.

Project description:White-rot fungi employ secreted carbohydrate-active enzymes (CAZymes) along with reactive oxygen species (ROS), like hydrogen peroxide (H2O2), to degrade lignocellulose in wood. H2O2 serves as a co-substrate for key oxidoreductases during the initial decay phase. While the degradation of lignocellulose by CAZymes is well documented, the impact of ROS on the oxidation of the secreted proteins remains unclear, and the identity of the oxidized proteins is unknown. Methionine (Met) can be oxidized to Met sulfoxide (MetO) or Met sulfone (MetO2) with potential deleterious, antioxidant, or regulatory effects. Other residues, like proline (Pro), can undergo carbonylation. Using the white-rot Pycnoporus cinnabarinus grown on aspen wood, we analyzed the Met content of the secreted proteins and their susceptibility to oxidation combining H218O2 with deep shotgun proteomics. Strikingly, their overall Met content was significantly lower (1.4%) compared to intracellular proteins (2.1%), a feature conserved in fungi but not in metazoans or plants. We evidenced that a catalase, widespread in white-rot fungi, protects the secreted proteins from oxidation. Our redox proteomics approach allowed the identification of 49 oxidizable Met and 40 oxidizable Pro residues within few secreted proteins, mostly CAZymes. Interestingly, many of them had several oxidized residues localized in hotspots. Some Met, including those in GH7 cellobiohydrolases, were oxidized up to 47%, with a substantial percentage of sulfone (13%). These Met are conserved in fungal homologs, suggesting important functional roles. Our findings reveal that white-rot fungi safeguard their secreted proteins by minimizing their Met content and by scavenging ROS and pinpoint redox-active residues in CAZymes.IMPORTANCEThe study of lignocellulose degradation by fungi is critical for understanding the ecological and industrial implications of wood decay. While carbohydrate-active enzymes (CAZymes) play a well-established role in lignocellulose degradation, the impact of hydrogen peroxide (H2O2) on secreted proteins remains unclear. This study aims at evaluating the effect of H2O2 on secreted proteins, focusing on the oxidation of methionine (Met). Using the model white-rot fungi Pycnoporus cinnabarinus grown on aspen wood, we showed that fungi protect their secreted proteins from oxidation by reducing their Met content and utilizing a secreted catalase to scavenge exogenous H2O2. The research identified key oxidizable Met within secreted CAZymes. Importantly, some Met, like those of GH7 cellobiohydrolases, undergone substantial oxidation levels suggesting important roles in lignocellulose degradation. These findings highlight the adaptive mechanisms employed by white-rot fungi to safeguard their secreted proteins during wood decay and emphasize the importance of these processes in lignocellulose breakdown.

Dataset Information

Bacteriophage-encoded virion-associated enzymes to overcome the carbohydrate barriers during the infection process.

Publications

Bacteriophage-encoded virion-associated enzymes to overcome the carbohydrate barriers during the infection process.

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