Cellular metabolism of unnatural sialic acid precursors

Pham, Nam D.; Fermaintt, Charles S.; Rodríguez, A. Carlos Blesa; McCombs, Janet E.; Nischan, Nicole; Kohler, Jennifer J.

doi:10.1007/s10719-015-9593-7

Cited by 28 publications

(53 citation statements)

References 44 publications

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“…Sarcoma 180 cell line was cultured in RPMI 1640 medium [ 20 ] containing 10% fetal bovine serum, 200 U/ml penicillin, and 200 μg/ml streptomycin at 37 °C, 5% CO 2 in humidified incubator.…”

Section: Methodsmentioning

confidence: 99%

Evaluating the antimicrobial, apoptotic, and cancer cell gene delivery properties of protein-capped gold nanoparticles synthesized from the edible mycorrhizal fungus Tricholoma crassum

et al. 2018

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Biosynthesis of gold nanoparticles of distinct geometric shapes with highly functional protein coats without additional capping steps is rarely reported. This study describes green synthesis of protein-coated gold nanoparticles for the first time from the edible, mycorrhizal fungus Tricholoma crassum (Berk.) Sacc. The nanoparticles were of the size range 5–25 nm and of different shapes. Spectroscopic analysis showed red shift of the absorption maxima with longer reaction period during production and blue shift with increase in pH. These were characterized with spectroscopy, SEM, TEM, AFM, XRD, and DLS. The particle size could be altered by changing synthesis parameters. These had potent antimicrobial activity against bacteria, fungi, and multi-drug-resistant pathogenic bacteria. These also had inhibitory effect on the growth kinetics of bacteria and germination of fungal spores. These showed apoptotic properties on eukaryotic cells when tested with comet assays. Moreover, the particles are capped with a natural 40 kDa protein which was utilized as attachment sites for genes to be delivered into sarcoma cancer cells. The present work also attempted at optimizing safe dosage of these nanoparticles using hemolysis assays, for application in therapy. Large-scale production of the nanoparticles in fermentors and other possible applications of the particles have been discussed.Electronic supplementary materialThe online version of this article (10.1186/s11671-018-2561-y) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Evaluating the antimicrobial, apoptotic, and cancer cell gene delivery properties of protein-capped gold nanoparticles synthesized from the edible mycorrhizal fungus Tricholoma crassum

et al. 2018

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“…Although mannosamine derivatives are relatively well tolerated by many cell lines (Almaraz et al 2012;Keppler et al 2001;Pham et al 2015), high concentrations may be cytotoxic for susceptible cell types such as neurons (Almaraz et al 2012;Du et al 2009). Mature (day 5, d5) 1 3 LUHMES cells were exposed to increasing concentration of the peracetylated sugars Ac 4 ManNAc and Ac 4 ManNAz for 24 h. Both, the neurite integrity and number of viable cells was measured and concentrations of ≥ 20 µM were found to be cytotoxic (Fig.…”

Section: Use Of Mge For Incorporation Of Labeled Sia Into Cell Surfacmentioning

confidence: 99%

Time and space-resolved quantification of plasma membrane sialylation for measurements of cell function and neurotoxicity

et al. 2019

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While there are many methods to quantify the synthesis, localization, and pool sizes of proteins and DNA during physiological responses and toxicological stress, only few approaches allow following the fate of carbohydrates. One of them is metabolic glycoengineering (MGE), which makes use of chemically modified sugars (CMS) that enter the cellular biosynthesis pathways leading to glycoproteins and glycolipids. The CMS can subsequently be coupled (via bio-orthogonal chemical reactions) to tags that are quantifiable by microscopic imaging. We asked here, whether MGE can be used in a quantitative and time-resolved way to study neuronal glycoprotein synthesis and its impairment. We focused on the detection of sialic acid (Sia), by feeding human neurons the biosynthetic precursor N-acetyl-mannosamine, modified by an azide tag. Using this system, we identified non-toxic conditions that allowed live cell labeling with high spatial and temporal resolution, as well as the quantification of cell surface Sia. Using combinations of immunostaining, chromatography, and western blotting, we quantified the percentage of cellular label incorporation and effects on glycoproteins such as polysialylated neural cell adhesion molecule. A specific imaging algorithm was used to quantify Sia incorporation into neuronal projections, as potential measure of complex cell function in toxicological studies. When various toxicants were studied, we identified a subgroup (mitochondrial respiration inhibitors) that affected neurite glycan levels several hours before any other viability parameter was affected. The MGE-based neurotoxicity assay, thus allowed the identification of subtle impairments of neurochemical function with very high sensitivity.

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“…used MOE to introduce azide‐containing glycoproteins into cell surface using N ‐azidoacetylmannosamine (ManNAz) as a biosynthetic precursor . Based on the studies of Kohler and others, the metabolic efficiency of different unnatural sialic acid precursors was cell line‐dependent for promiscuous intracellular environments . Some experiments demonstrated that K562, SH‐SY, HeLa and PC‐3 cells were almost unable to metabolize some unnatural modified mannosamine analogs to membrane sialic acid glycoconjugates.…”

Section: Tools To Probe Siglecsmentioning

confidence: 99%

Probing Sialidases or Siglecs with Sialic Acid Analogues, Clusters and Precursors

2019

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Sialic acids have recently gathered interests of many researchers. Sialidases and Siglecs are two important sialic acids-related proteins that exist in biological systems, and aberrant sialoside-Siglec or sialoside-sialidase interactions are associated with many diseases, including autoimmunity, infection and cancer. Probing sialidase and Siglec effectively can enable the studies on the complex physiological functions of sialic acids. Over the last few decades, many sialic acid derivatives with modifications of parent skeleton and aglycon have been designed to increase the binding affinity with sialidase/Siglec or improve their detection efficiency. At the same time, in-situ detection has also been aided by metabolic oligosaccharide engineering, in which incubating cells with sialic acid precursors could result in unnatural sialylated glycans incorporated into cellular membrane. In this minireview, we will predominantly highlight the development of sialidase and Siglec probing strategies using sialic acid analogues, clusters and precursors, which may further lay foundation for new diagnostics and treatments. 2 3 4 5 6 7 8

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Cellular metabolism of unnatural sialic acid precursors

Cited by 28 publications

References 44 publications

Evaluating the antimicrobial, apoptotic, and cancer cell gene delivery properties of protein-capped gold nanoparticles synthesized from the edible mycorrhizal fungus Tricholoma crassum

Evaluating the antimicrobial, apoptotic, and cancer cell gene delivery properties of protein-capped gold nanoparticles synthesized from the edible mycorrhizal fungus Tricholoma crassum

Time and space-resolved quantification of plasma membrane sialylation for measurements of cell function and neurotoxicity

Probing Sialidases or Siglecs with Sialic Acid Analogues, Clusters and Precursors

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