Carbohydrate Modification Research Articles

Photocatalyst-free light-promoted carbohydrate synthesis and modification

Photoredox catalysis has recently emerged as a powerful approach for preparing oligosaccharides because it uses mild conditions, is compatible with partially or completely unprotected carbohydrate substrates, and exhibits impressive regio‐ and stereo‐selectivity and high functional group tolerance. However, most catalytic photoredox reactions require an external photocatalyst (organic dye or expensive transition-metal complex) to deliver key glycosyl radicals. Several photocatalyst-free photocatalytic reactions that avoid the use of expensive metal salts or organic-dye additives have received significant attention. In this review, we highlight the most recent developments in photocatalyst-free light-promoted carbohydrate synthesis and modification, which is expected to inspire broad interest in further innovations in the green synthesis of saccharides.

Clinical glycoprotein mass spectrometry: The future of disease detection and monitoring.

Protein glycosylation is the co- and/or post-translational modification of proteins with oligosaccharides (glycans). This process is not template based and can introduce a heterogeneous set of glycan modifications onto substrate proteins. Glycan structures preserve biomolecular information from the cell, with glycoproteins from different cell types and tissues displaying distinct patterns of glycosylation. Several decades of research have revealed that glycan structures also differ between normal physiology and disease. This suggests that the information stored in glycoproteins and glycans can be utilized for disease diagnosis and monitoring. Methods that enable sensitive and site-specific measurement of protein glycosylation in clinical settings, such as nano-flow liquid chromatography tandem mass spectrometry, are therefore essential. The purpose of this perspective is to discuss recent advances in mass spectrometry and the potential of these advances to facilitate the detection and monitoring of disease-specific glycoprotein glycoforms. Glycoproteomics, the system-wide characterization of glycoprotein identity inclusive of site-specific characterization of carbohydrate modifications on proteins, and glycomics, the characterization of glycan structures, will be discussed in this context. Quantitative measurement of glycopeptide markers via parallel reaction monitoring is highlighted. The development of promising glycopeptide markers for autoimmune disease, liver disease, and liver cancer is discussed. Synthetic glycopeptide standards, ambient ionization mass spectrometry, and consideration of glyco-biomarkers in two- and three-dimensional space within tissue will be critical to the advancement of this field. The authors envision a future in which glycoprotein mass spectrometry workflows will be integrated into clinical settings, to aid in the rapid diagnosis and monitoring of disease.

Mini-Review: The Distinct Carbohydrate Active Enzyme Secretome of Rhizopus spp. Represents Fitness for Mycelium Remodeling and Solid-State Plant Food Fermentation.

Rhizopus is a genus of filamentous fungi belonging to the Mucoromycota division. Rhizopus species produce a white, dense mycelium, which is used to create tempeh, a solid-state fermented Asian soybean product, that is gaining renewed attention as a proteinaceous plant food. The profile of carbohydrate-active enzymes (CAZymes) of a fungus or group of fungi, particularly the secretome CAZymes profile, reflects adaptation to different lifestyles and habitats, and has a significant impact on fermentative capacity. This review examines the CAZymes profiles of Rhizopus species focusing on their implication for carbohydrate utilization and solid-state fermentation of plant materials. Through comprehensive genomic assessments and comparisons with other filamentous fungi, we particularly highlight how the unique CAZymes secretome profile is closely correlated with the taxonomy and ecological niches of Rhizopus species. We discuss how the CAZymes secretome capacity of Rhizopus species differs from other fungi and summarize the current state of knowledge regarding the specific CAZymes involved in the modification of carbohydrates in the fungal cell wall and in plant cell walls. We foresee that advanced genomic and proteomic technologies will be used to expand the biotechnology applications of Rhizopus spp.

Stereoselective Construction of Multifunctional C-Glycosides Enabled by Nickel-Catalyzed Tandem Borylation/Glycosylation.

Stereochemically pure saccharides have indispensable roles in fields ranging from medicinal chemistry to materials science and organic synthesis. However, the development of a simple, stereoselective, and efficient glycosylation protocol to access α- and β-C-glycosides (particularly 2-deoxy entities) remains a persistent challenge. Existing studies have primarily focused on C1 modification of carbohydrates and transformation of glycosyl radical precursors. Here, we innovate by harnessing the in situ generated glycosyl-Ni species to achieve one-pot borylation and glycosylation in a cascade manner, which is enabled by an earth-abundant nickel-catalyzed carboboration of readily accessible glycals without any ligand. This work reveals the potential for the development of a modular and multifunctional glycosylation platform to facilitate the simultaneous introduction of C-C and C-B bonds at the stereogenic center of saccharides, a largely unexploited research area. Preliminary experimental and computational studies indicate that the endocyclic O and the C3 group play important roles in stereoseclectively forging glycosidic bonds. As a result, a diverse range of C-R (R = alkyl, aryl, and alkenyl) and 2-deoxygenated glycosides bearing modifiable boron groups could be rapidly made with excellent stereocontrol and exhibit remarkable functional group tolerance. The synthetic potential is underscored in the late-stage glycosylation of natural products and commercial drugs as well as the facile preparation of various rare sugars, bioactive conjugates, and key intermediates to prorocentin, phomonol, and aspergillide A.

Stereoselective and controllable C3-Amination or C1,C3-di-amination of 2-nitroglycals

Precise and selective modification of carbohydrates is a critical strategy in producing diverse carbohydrate derivatives for exploiting their functions. We disclosed a simple, efficient, and highly regioselective and stereoselective protocol to controllable amination of 2-nitroglycals under mild conditions in 5 min. A range of 3-amino-carbohydrates including 3-arylamino-2-nitro-glycals and 1,3-di-amino-carbohydrate derivatives were obtained in good to excellent yield with excellent stereoselectivity. The produced 3-amino-2-nitro-glycals can be used as a precursor for further transformation.

Abstract 2984: Prostate cancer associated glycosylated RNAs: Who are you and where are you hiding

Abstract Glycosylated RNA (glycoRNA) is an emerging field. Carbohydrate modifications are vital for biomolecule (proteins and lipids) stability, transport, and localization. Glycosylation influences cell functions like differentiation, apoptosis, and immune response, potentially impacting human disease. The Bertozzi lab initially discovered that RNAs can carry sugar modifications when they metabolically labeled mammalian cultured cells using bioorthogonal chemistry methods normally employed for glycosylated protein and lipid enrichment. Our lab wanted to investigate the translational potential of glycoRNAs and determine if these modifications could be developed as novel clinical targets for aggressive prostate cancer. (PCa). As advanced forms of PCa often lead to poor prognoses, it is imperative that better screening biomarkers and therapeutic treatment options are developed to increase patient survivorship. We predicted that glycoRNA expression would correlate with malignancy, metastatic status, and hormone sensitivity in prostate cells. Our lab used the sialic acid metabolic precursor Ac4ManNAz and click chemistry to label a panel of human prostate cancer cell lines representing the spectrum of prostate disease followed by rigorous RNA isolation procedures and northern blot analysis. We found that human prostate cell lines with no (RWPE-1) or low malignancy (LNCaP) expressed higher levels of glycoRNA belonging to the small <200 nt (but not large) noncoding RNA class compared to aggressive, metastatic PCa cell lines. Our glycosidase studies and lectin northern blot analysis indicated that RNA carries both N-linked and O-linked modifications. Based on these findings, we hypothesized that glycosylated RNA plays an important role in maintaining prostate homeostasis and in cancer protection. To better profile these glycoRNAs, we aimed to: 1) Test a wider range of metabolic labeling regents to characterize the glycoRNA populations using a panel of human prostate cell lines; 2) Perform cellular fractionation studies to determine localization of glycoRNA in prostate cells; and 3) Explore glycoRNAs in a genetic animal model. These studies focused on LNCaP PCa cell lines, which express high levels of glycoRNA. We found that small glycoRNA expression was noted when LNCaP cells were metabolically labeled with Ac4GalNAz (predominantly O-linked) and Ac4ManNAz (predominantly N-linked), verifying glycoRNA heterogeneity. Cellular fractionation studies indicated glycoRNA localization was enriched in the membrane fraction of LNCaP cells using these metabolic labels. Furthermore, we verified that C. elegans grown on Ac4GalNAz plates expressed small glycoRNA, indicating biological conservation. We are now characterizing these glycoRNAs using RNA sequencing and LC-MS/MS glycomics. Our findings will provide novel insights into cancer-associated glycoRNA, leading to improved clinical tools for PCa. Citation Format: Samantha McGuire, Esther Jones, Spencer Moen, Aurora Esquela-Kerscher. Prostate cancer associated glycosylated RNAs: Who are you and where are you hiding [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2984.

Controllable 1,3-Bis-Functionalization of 2-Nitroglycals with High Regioselectivity and Stereoselectivity Enabled by a H-Bond Catalyst.

The selective modification of carbohydrates is significant for producing their unnatural analogues for drug discovery. C1-functionalization (glycosylation) and C1,C2-difunctionalization of carbohydrates have been well developed. In contrast, C3-functionalization or C1,C3-difunctionalization of carbohydrates remains rare. Herein, we report such processes that efficiently and stereoselectively modify carbohydrates. Specifically, we found that trifluoroethanol (TFE) could promote 1,3-bis-indolylation/pyrrolylation of 2-nitroglycals generated carbohydrate derivatives in up to 93% yield at room temperature; slightly reducing the temperature could install two different indoles at the C1- and C3-positions. Switching TFE to a bifunctional amino thiourea catalyst leads to the generation of C3 monosubstituted carbohydrates, which could also be used to construct 1,3-di-C-functionalized carbohydrates. This approach produced a range of challenging sugar derivatives (over 80 examples) with controllable and high stereoselectivity (single isomer for over 90% of the examples). The potential applications of the reaction were demonstrated by a set of transformations including the synthesis of bridged large-ring molecules and gram scale reactions. Biological activities evaluation demonstrated that three compounds exhibit a potent inhibitory effect on human cancer cells T24, HCT116, AGS, and MKN-45 with IC50 ranged from 0.695 to 3.548 μM.

Transcriptomic and biochemical insights into fall armyworm (Spodoptera frugiperda) responses on silicon-treated maize.

The fall armyworm, Spodoptera frugiperda, is an agricultural pest of significant economic concern globally, known for its adaptability, pesticide resistance, and damage to key crops such as maize. Conventional chemical pesticides pose challenges, including the development of resistance and environmental pollution. The study aims to investigate an alternative solution: the application of soluble silicon (Si) sources to enhance plant resistance against the fall armyworm. Silicon dioxide (SiO2) and potassium silicate (K2SiO3) were applied to maize plants via foliar spray. Transcriptomic and biochemical analyses were performed to study the gene expression changes in the fall armyworm feeding on Si-treated maize. Results indicated a significant impact on gene expression, with a large number of differentially expressed genes (DEGs) identified in both SiO2 and K2SiO3 treatments. Furthermore, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis identified critical DEGs involved in specific pathways, including amino acid, carbohydrate, lipid, energy, xenobiotics metabolisms, signal transduction, and posttranslational modification, significantly altered at both Si sources. Biochemical analyses further revealed that Si treatments inhibited several enzyme activities (glutamate dehydrogenase, trehalase, glucose-6-phosphate dehydrogenase, chitinase, juvenile hormone esterase, and cyclooxygenase while simultaneously inducing others (total protein, lipopolysaccharide, fatty acid synthase, ATPase, and cytochrome P450), thus suggesting a toxic effect on the fall armyworm. In conclusion, Si applications on maize influence the gene expression and biochemical activities of the fall armyworm, potentially offering a sustainable pest management strategy.

Carbohydrate-active enzyme (CAZyme) discovery and engineering via (Ultra)high-throughput screening.

Carbohydrate-active enzymes (CAZymes) constitute a diverse set of enzymes that catalyze the assembly, degradation, and modification of carbohydrates. These enzymes have been fashioned into potent, selective catalysts by millennia of evolution, and yet are also highly adaptable and readily evolved in the laboratory. To identify and engineer CAZymes for different purposes, (ultra)high-throughput screening campaigns have been frequently utilized with great success. This review provides an overview of the different approaches taken in screening for CAZymes and how mechanistic understandings of CAZymes can enable new approaches to screening. Within, we also cover how cutting-edge techniques such as microfluidics, advances in computational approaches and synthetic biology, as well as novel assay designs are leading the field towards more informative and effective screening approaches.

An Arabidopsis mutant line lacking the mitochondrial calcium transport regulator MICU shows an altered metabolite profile

ABSTRACT Plant metabolism is constantly changing and requires input signals for efficient regulation. The mitochondrial calcium uniporter (MCU) couples organellar and cytoplasmic calcium oscillations leading to oxidative metabolism regulation in a vast array of species. In Arabidopsis thaliana, genetic deletion of AtMICU leads to altered mitochondrial calcium handling and ultrastructure. Here we aimed to further assess the consequences upon genetic deletion of AtMICU. Our results confirm that AtMICU safeguards intracellular calcium transport associated with carbohydrate, amino acid, and phytol metabolism modifications. The implications of such alterations are discussed.

Practical Site-selective Oxidation of Glycosides with Pd(OAc)2/Neocuproine

Palladium-catalyzed oxidation of the secondary C3 hydroxy group in glycopyranosides has set its mark in the selective modification of unprotected carbohydrates. The pre-formed catalyst [(neocuproine)PdOAc]2(OTf)2 oxidizes besides monosaccharides also di-, and oligosaccharides. Here, we provide a more convenient protocol for this reaction in which the Pd-catalyst is formed in situ from Pd(OAc)2 and neocuproine in methanol at 50 °C. Together with a simplified product isolation, this protocol is applied to a series of mono- and disaccharides and has been applied on 10-gram scale. The protocol is also valuable as a screening method to determine whether more extensive studies using the pre-formed catalyst are worthwhile.

Polysaccharide metabolism in Anacardiaceae (Asian lacquer) cross-linked polymers elucidated using in situ trimethylsilylation pyrolysis-gas chromatography–mass spectrometry

Carbohydrates from polysaccharides in natural thermoset Anacardiaceae polymers of Gluta usitata, Toxicodendron succedaneum and Toxicodendron vernicifluum were identified using pyrolysis-gas chromatography–mass spectrometry with in situ trimethylsilylation. Pyrolysates resulting from the pyrolytic intermolecular chain scission of the polysaccharides were used to elucidate monomeric units. Polysaccharides, dispersed in the phenolic lacasse catalysed cross-linked macromolecules, showed to be metabolised through various catabolic and anabolic routes. Galactose functionalities, abundantly present in the polysaccharides were determined to be enzymatically converted to glucose-6-phosphate, followed by conversion via glycolysis and pentose phosphate pathways. Determination of specific routes of carbohydrate modification via glycolysis and pentose phosphate pathways facilitated differentiating G. usitata, T. succedaneum and T. vernicifluum polymers, based on the carbohydrate content. It was also found that uronic type acids, present as end groups of the branched polysaccharide structure, were biochemically converted to aldonic acids. Following the pentose phosphate and glycolysis routes, carbohydrates in G. usitata and T. vernicifluum polymers showed to be further modified via shikimate and cinnamate pathways to produce phenylpropanoid compounds. Parent molecules and pyrolysis products thereof were verified using analytical standards of high purity. The mass spectra and Kovats retention indices were compiled in an AMDIS library, which can be made available on request.

A mini review on manipulation of carbohydrate for better use in surimi and surimi products: modification and compounding.

Carbohydrate is widely used in the production of surimi and surimi products to improve their qualities, such as anti-freezing capability, gelling ability, nutrition, flavor and 3D printability. More and more native carbohydrates have been modified through physical methods (e.g., ball milling, irradiation and differential sedimentation), chemical method (e.g., deacetylation, hydroxypropylation and acetic acid esterification) or enzymatic method (e.g., chitosanase) before being used in the processing of surimi and surimi products in recent years. At the same time, different carbohydrates are compounded and applied to surimi and surimi products. The modified and compounded carbohydrates in surimi have been proved to improve quality of surimi and surimi products more pronouncedly than native carbohydrates. Therefore, this review summarizes the manipulation of carbohydrate by modification and compounding to improve the qualities of surimi and surimi products. Moreover, the prospects for carbohydrate modification and compounding for use in surimi and surimi products are discussed. © 2023 Society of Chemical Industry.

How to Develop Drug Delivery System Based on Carbohydrate Nanoparticles Targeted to Brain Tumors.

Brain tumors are the most difficult to treat, not only because of the variety of their forms and the small number of effective chemotherapeutic agents capable of suppressing tumor cells, but also limited by poor drug transport across the blood-brain barrier (BBB). Nanoparticles are promising drug delivery solutions promoted by the expansion of nanotechnology, emerging in the creation and practical use of materials in the range from 1 to 500 nm. Carbohydrate-based nanoparticles is a unique platform for active molecular transport and targeted drug delivery, providing biocompatibility, biodegradability, and a reduction in toxic side effects. However, the design and fabrication of biopolymer colloidal nanomaterials have been and remain highly challenging to date. Our review is devoted to the description of carbohydrate nanoparticle synthesis and modification, with a brief overview of the biological and promising clinical outcomes. We also expect this manuscript to highlight the great potential of carbohydrate nanocarriers for drug delivery and targeted treatment of gliomas of various grades and glioblastomas, as the most aggressive of brain tumors.

Small Molecule Inhibition of an Exopolysaccharide Modification Enzyme is a Viable Strategy To Block Pseudomonas aeruginosa Pel Biofilm Formation.

Biosynthesis of the Pel exopolysaccharide in Pseudomonas aeruginosa requires all seven genes of the pelABCDEFG operon. The periplasmic modification enzyme PelA contains a C-terminal deacetylase domain that is necessary for Pel-dependent biofilm formation. Herein, we show that extracellular Pel is not produced by a P. aeruginosa PelA deacetylase mutant. This positions PelA deacetylase activity as an attractive target to prevent Pel-dependent biofilm formation. Using a high-throughput screen (n = 69,360), we identified 56 compounds that potentially inhibit PelA esterase activity, the first enzymatic step in the deacetylase reaction. A secondary biofilm inhibition assay identified methyl 2-(2-pyridinylmethylene) hydrazinecarbodithioate (SK-017154-O) as a specific Pel-dependent biofilm inhibitor. Structure-activity relationship studies identified the thiocarbazate as a necessary functional group and that the pyridyl ring could be replaced with a phenyl substituent (compound 1). Both SK-017154-O and compound 1 inhibit Pel-dependent biofilm formation in Bacillus cereus ATCC 10987, which has a predicted extracellular PelA deacetylase in its pel operon. Michaelis-Menten kinetics determined SK-017154-O to be a noncompetitive inhibitor of PelA, while compound 1 did not directly inhibit PelA esterase activity. Cytotoxicity assays using human lung fibroblast cells showed that compound 1 is less cytotoxic than SK-017154-O. This work provides proof of concept that biofilm exopolysaccharide modification enzymes are important for biofilm formation and can serve as useful antibiofilm targets. IMPORTANCE Present in more than 500 diverse Gram-negative and 900 Gram-positive organisms, the Pel polysaccharide is one of the most phylogenetically widespread biofilm matrix determinants found to date. Partial de-N-acetylation of this α-1,4 linked N-acetylgalactosamine polymer by the carbohydrate modification enzyme PelA is required for Pel-dependent biofilm formation in Pseudomonas aeruginosa and Bacillus cereus. Given this and our observation that extracellular Pel is not produced by a P. aeruginosa PelA deactylase mutant, we developed an enzyme-based high-throughput screen and identified methyl 2-(2-pyridinylmethylene) hydrazinecarbodithioate (SK-017154-O) and its phenyl derivative as specific Pel-dependent biofilm inhibitors. Michaelis-Menten kinetics revealed SK-017154-O is a noncompetitive inhibitor and that its noncytotoxic, phenyl derivative does not directly inhibit P. aeruginosa PelA esterase activity. We provide proof of concept that exopolysaccharide modification enzymes can be targeted with small molecule inhibitors to block Pel-dependent biofilm development in both Gram-negative and Gram-positive bacteria.

The efficacy of a home-use metabolic device (Lumen) in response to a short-term low and high carbohydrate diet in healthy volunteers.

Based on stoichiometric assumptions, and real-time assessment of expired carbon dioxide (%CO2) and flow rate, the Lumen device provides potential for consumers/athletes to monitor metabolic responses to dietary programs outside of laboratory conditions. However, there is a paucity of research exploring device efficacy. This study aimed to evaluate Lumen device response to: i) a high-carbohydrate meal under laboratory conditions, and ii) a short-term low- or high-carbohydrate diet in healthy volunteers. Following institutional ethical approval, 12 healthy volunteers (age: 36±4yrs; body mass: 72.1±3.6 kg; height: 1.71±0.02 m) performed Lumen breath and Douglas bag expired air measures under fasted laboratory conditions and at 30 and 60min after a high-carbohydrate (2g·kg-1) meal, along with capilliarized blood glucose assessment. Data were analyzed using a one-way ANOVA, with ordinary least squares regression used to assess the model between Lumen expired carbon dioxide percentage (L%CO2) and respiratory exchange ratio (RER). In a separate phase, 27 recreationally active adults (age: 42±2yrs; body mass: 71.9±1.9 kg; height: 1.72±0.02 m) completed a 7-day low- (~20% of energy intake [EI]; LOW) or high-carbohydrate diet (~60% of EI; HIGH) in a randomized, cross-over design under free-living conditions. L%CO2 and derived Lumen Index (LI) were recorded daily across morning (fasted and post-breakfast) and evening (pre/post meal, pre-bed) periods. Repeated measures ANOVA were employed for main analyses, with Bonferroni post-hoc assessment applied (P ≤0.05). Following the carbohydrate test-meal, L%CO2 increased from 4.49±0.05% to 4.80±0.06% by 30min, remaining elevated at 4.76±0.06% by 60min post-feeding (P <0.001, ηp2=0.74). Similarly, RER increased by 18.1% from 0.77±0.03 to 0.91±0.02 by 30min post-meal (P =0.002). When considering peak data, regression analysis demonstrated a significant model effect between RER and L%CO2 (F=5.62, P =0.03, R2=0.20). Following main dietary interventions, no significant interactions (diet × day) were found. However, main diet effects were evident across all time-points assessed, highlighting significant differences for both L%CO2 and LI between LOW and HIGH conditions (P <0.003). For L%CO2, this was particularly noted under fasted (4.35±0.07 vs. 4.46±0.06%, P =0.001), pre-evening meal (4.35±0.07 vs. 4.50±0.06%, P <0.001), and pre-bed time-points (4.51±0.08 vs. 4.61±0.06%, P =0.005). Our findings demonstrated that a portable, home-use metabolic device (Lumen) detected significantly increased expired %CO2 in response to a high-carbohydrate meal, and may be useful in tracking mean weekly changes to acute dietary carbohydrate modifications. Additional research is warranted to further determine the practical and clinical efficacy of the Lumen device in applied compared to laboratory settings.

Mixing nitrogenous ligands in osmium(VI)ligand-polysaccharide complexes. Voltammetric sensing

• Electroinactive dextran was modified with complexes of Os(VI) and nitrogenous ligands. • Os(VI)ligand modified dextran is electroactive and yields several redox peaks. • Nitrogenous ligand type significantly influences interfacial properties of modified dextrans. • Ligand combination can improve dextran analysis comparing to one-ligand modification. Simple chemical modification of carbohydrates with complexes of sixvalent osmium and nitrogenous ligands (Os(VI)L) changes electro inactive carbohydrates to electroactive species. In this work we show that the type of the nitrogenous ligand (L) significantly influences adsorption of Os(VI)L-modified dextrans at a charged surface. Modification by mixture containing two ligands can improve electrochemical analysis in comparison to simple modification only with one ligand. For instance, modification with complexes of Os(VI) and mixture of N,N,Ń,Ń-tetramethylethylenediamine (temed) with 2,2́-bipyridine (bipy) allows to (i) determine the concentration of Os(VI)L modified dextran in one order higher range as in case of dextran modified only with Os(VI)temed and (ii) discriminate between various polysaccharides differing in building units and structures.

Deletion of POMT2 in Zebrafish Causes Degeneration of Photoreceptors.

Mutations in the extracellular matrix protein eyes shut homolog (EYS) are a common cause of retinitis pigmentosa, a blinding disease characterized by photoreceptor degeneration. EYS binds to matriglycan, a carbohydrate modification on O-mannosyl glycan substitutions of the cell-surface glycoprotein α-dystroglycan. Patients with mutations in enzymes required for the biosynthesis of matriglycan exhibit syndromic retinal atrophy, along with brain malformations and congenital muscular dystrophy. Protein O-mannosyltransferase 2 (POMT2) is an enzyme required for the synthesis of O-mannosyl glycans. To evaluate the roles of O-mannosyl glycans in photoreceptor health, we generated protein O-mannosyltransferase 2 (pomt2) mutant zebrafish by CRISPR. pomt2 mutation resulted in a loss of matriglycan and abolished binding of EYS protein to α-dystroglycan. Mutant zebrafish presented with hydrocephalus and hypoplasia of the cerebellum, as well as muscular dystrophy. EYS protein was enriched near photoreceptor connecting cilia in the wild-type, but its presence and proper localization was significantly reduced in mutant animals. The mutant retina exhibited mis-localization of opsins and increased apoptosis in both rod and cone photoreceptors. Immunofluorescence intensity of G protein subunit alpha transducin 2 (GNAT2) antibody (a general cone marker) and 1D4 antibody (a long double cone marker) in mutant retinas did not differ from wild-type retinas at 1-month post fertilization, but was reduced at 6 months post fertilization, indicating significant cone degeneration. These data suggest that POMT2-mediated O-mannosyl glycosylation is required for EYS protein localization to the connecting cilium region and photoreceptor survival.

Recent Advances in the Development and Synthesis of Carbohydrate‐Based Molecules with Promising Antibacterial Activity

AbstractWith the rise of antimicrobial resistance (AMR), innovation in antibacterial drug research and development is urgently needed and strongly encouraged by the World Health Organization (WHO). Carbohydrates are valuable bioactive scaffolds to be explored in this context, and because of their unique multifunctionality, stereochemical diversity, and natural protein‐binding profile, they come across as attractive starting materials for the synthesis of antimicrobial agents with innovative mechanisms of action (MoA). In this concise review, state‐of‐the‐art methodologies for the synthesis of an array of promising and recently developed carbohydrate‐based molecules with antibacterial activity are presented and discussed. By describing successful case studies as platforms for the scrutiny of carbohydrate modification and coupling approaches in organic chemistry, this work summarizes the latest research efforts in this area, ultimately encouraging the design and synthesis of new and much‐needed glycoantibiotic leads for pharmaceutical development.

Exploiting photoredox catalysis for carbohydrate modification through C-H and C-C bond activation.

Photoredox catalysis has recently emerged as a powerful synthetic platform for accessing complex chemical structures through non-traditional bond disconnection strategies that proceed through free-radical intermediates. Such synthetic strategies have been used for a range of organic transformations; however, in carbohydrate chemistry they have primarily been applied to the generation of oxocarbenium ion intermediates in the ubiquitous glycosylation reaction. In this Review, we present more intricate light-induced synthetic strategies to modify native carbohydrates through homolytic C-H and C-C bond cleavage. These strategies allow access to glycans and glycoconjugates with profoundly altered carbohydrate skeletons, which are challenging to obtain through conventional synthetic means. Carbohydrate derivatives with such structural motifs represent a broad class of natural products integral to numerous biochemical processes and can be found in active pharmaceutical substances. Here we present progress made in C-H and C-C bond activation of carbohydrates through photoredox catalysis, focusing on the operational mechanisms and the scope of the described methodologies.