Metabolism of Vertebrate Amino Sugars with N-Glycolyl Groups

Macauley, Matthew S.; Chan, Jefferson; Zandberg, Wesley F.; He, Yuan; Whitworth, Garrett E.; Stubbs, Keith A.; Yuzwa, Scott A.; Bennet, Andrew J.; Varki, Ajit; Davies, G.J.; Vocadlo, David J.

doi:10.1074/jbc.m112.363721

Cited by 25 publications

(11 citation statements)

References 56 publications

(49 reference statements)

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“…3C). Kinetic parameters for this reaction were similar to reported values, 2627 confirming the reliability of the continuous assay measurements. Both the C215A and C215G mutants retained activity toward p NP-GlcNAc, displaying catalytic efficiencies ( k cat /K m ) 2- to 3-fold lower than the wild-type enzyme.…”

Section: Resultssupporting

confidence: 84%

Recognition of diazirine-modified O-GlcNAc by human O-GlcNAcase

Rodríguez

Kohler

2014

Med. Chem. Commun.

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The mammalian O-GlcNAc hydrolase (OGA) removes O-GlcNAc from serine and threonine residues on intracellular glycoproteins. OGA activity is sensitive to N-acyl substitutions to O-GlcNAc, with alkyl diazirine-modified O-GlcNAc (O-GlcNDAz) being completely resistant to removal by OGA. Using homology modeling, we identified OGA residues proximal to the N-acyl position of O-GlcNAc substrate. Mutation of one of these residues, C215, results in mutant enzymes that are able to hydrolytically remove O-GlcNDAz from a model compound. Further, the C215A mutant is capable of removing O-GlcNDAz from a peptide substrate. These results can be used to improve metabolism of O-GlcNAc analogs in cells. In addition, the enzyme specificity studies reported here provide new insight into the active site of OGA, an important drug target.

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Section: Resultssupporting

confidence: 84%

Recognition of diazirine-modified O-GlcNAc by human O-GlcNAcase

Rodríguez

Kohler

2014

Med. Chem. Commun.

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“…By applying Zemplen conditions, fully protected intermediate 2 was next deprotected, and GlcNGc was achieved in good yield. Spectral properties of the desired products compounds 2 , and GlcNGc matched those described previously (Sinaÿ 1971 ; Bergfeld et al 2012 ; Macauley et al 2012 ).…”

Section: Methodssupporting

confidence: 75%

N-glycolyl chondroitin synthesis using metabolically engineered E. coli

et al. 2020

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N-glycolyl chondroitin (Gc-CN) is a metabolite of N-glycolylneuraminic acid (Neu5Gc), a sialic acid that is commonly found in mammals, but not humans. Humans can incorporate exogenous Neu5Gc into their tissues from eating red meat. Neu5Gc cannot be biosynthesized by humans due to an evolutionary mutation and has been implicated in causing inflammation causing human diseases, such as cancer. The study Neu5Gc is important in evolutionary biology and the development of potential cancer biomarkers. Unfortunately, there are several limitations to detecting Neu5Gc. The elimination of Neu5Gc involves a degradative pathway leading to the incorporation of N-glycolyl groups into glycosaminoglycans (GAGs), such as Gc-CN. Gc-CN has been found in humans and in animals including mice, lamb and chimpanzees. Here, we present the biosynthesis of Gc-CN in bacteria by feeding chemically synthesized N-glycolylglucosamine to Escherichia coli. A metabolically engineered strain of E. coli K4, fed with glucose supplemented with GlcNGc, converted it to N-glycolylgalactosamine (GalNGc) that could then be utilized as a substrate in the chondroitin biosynthetic pathway. The final product, Gc-CN was converted to disaccharides using chondroitin lyase ABC and analyzed by liquid chromatography-tandem mass spectrometry with multiple reaction monitoring detection. This analysis showed the incorporation of GalNGc into the backbone of the chondroitin oligosaccharide.

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“…By applying Zemplen conditions, fully protected intermediate 2 was next deprotected, and GlcNGc was achieved in good yield. Spectral properties of the desired products compounds 2, and GlcNGc matched those described previously (Sinaÿ 1971;Bergfeld et al 2012;Macauley et al 2012).…”

Section: N-glycolyl Glucosamine (Glcngc)-synthesis and Chemical Charasupporting

confidence: 74%

N-glycolyl chondroitin synthesis using metabolically engineered E. coli

Awofiranye

Baytas

Xia

et al. 2020

Preprint

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N -glycolylneuraminic acid (Neu5Gc) is a sialic acid missing in humans but commonly found in mammals and is the proposed cause of “xenosialitis” in humans, a “xenoautoimmune” inflammatory reaction believed to result from humans consuming Neu5Gc from red meat that human cells incorporate into some cell types, but is also recognized as a foreign antigen. The production of antibodies against Neu5Gc-containing glycans causes inflammation that is hypothesized to be an aggravating factor to multiple diseases associated with red meat consumption, including cancers arising from epithelial cells lining hollow organs and atherosclerosis involving vascular endothelial cells. The small amounts of Neu5Gc involved is glycosidically-linked at the reducing end of glycans but is difficult to quantitate because of its location on endothelial and epithelial cells. A more stable recently discovered metabolite of Neu5Gc, N-glycolyl chondroitin (Gc-CN), is present in the serum of humans who eat red meat and is more easily isolated and analyzed. Here, we present the biosynthesis of Gc-CN by feeding chemically synthesized N-glycolylglucosamine to Escherichia coli. A metabolically engineered strain of E. coli K4, fed with glucose supplemented with GlcNGc, converted it to N-glycolylgalactosamine (GalNGc) that could then be utilized as a substrate in the chondroitin biosynthetic pathway. The final product, Gc-CN was converted to disaccharides using chondroitin lyase ABC and analyzed by liquid chromatography-tandem mass spectrometry with multiple reaction monitoring detection. This analysis showed the incorporation of GalNGc into the backbone of the chondroitin oligosaccharide.

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Metabolism of Vertebrate Amino Sugars with N-Glycolyl Groups

Cited by 25 publications

References 56 publications

Recognition of diazirine-modified O-GlcNAc by human O-GlcNAcase

Recognition of diazirine-modified O-GlcNAc by human O-GlcNAcase

N-glycolyl chondroitin synthesis using metabolically engineered E. coli

N-glycolyl chondroitin synthesis using metabolically engineered E. coli

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