Enzymatic synthesis of nucleobase-modified UDP-sugars: scope and limitations

Wagstaff, Ben A.; Rejzek, Martin; Pesnot, Thomas; Tedaldi, Lauren; Caputi, Lorenzo; O’Neill, Ellis C.; Benini, Stefano; Wagner, Gerd K.; Field, Robert A.

doi:10.1016/j.carres.2014.12.005

Cited by 22 publications

(17 citation statements)

References 28 publications

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“…The reaction reached completion when signals for CTP (−5.83 ppm, doublet, −10.75 ppm, doublet, −19.38 ppm, triplet) had diminished. At this point, CMP–Kdn was purified using strong anion exchange as outlined in Wagstaff et al ( 64 ). 1 H NMR, 31 P NMR, and MS values aligned well with previously described literature ( 21 , 65 ) ( Fig.…”

Section: Methodsmentioning

confidence: 99%

Identification of a Kdn biosynthesis pathway in the haptophyte Prymnesium parvum suggests widespread sialic acid biosynthesis among microalgae

Wagstaff

Rejzek

Field

2018

Journal of Biological Chemistry

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Sialic acids are a family of more than 50 structurally distinct acidic sugars on the surface of all vertebrate cells where they terminate glycan chains and are exposed to many interactions with the surrounding environment. In particular, sialic acids play important roles in cell–cell and host–pathogen interactions. The sialic acids or related nonulosonic acids have been observed in Deuterostome lineages, Eubacteria, and Archaea but are notably absent from plants. However, the structurally related C8 acidic sugar 3-deoxy-d-manno-2-octulosonic acid (Kdo) is present in Gram-negative bacteria and plants as a component of bacterial lipopolysaccharide and pectic rhamnogalacturonan II in the plant cell wall. Until recently, sialic acids were not thought to occur in algae, but as in plants, Kdo has been observed in algae. Here, we report the de novo biosynthesis of the deaminated sialic acid, 3-deoxy-d-glycero-d-galacto-2-nonulosonic acid (Kdn), in the toxin-producing microalga Prymnesium parvum. Using biochemical methods, we show that this alga contains CMP–Kdn and identified and recombinantly expressed the P. parvum genes encoding Kdn-9-P synthetase and CMP–Kdn synthetase enzymes that convert mannose-6-P to CMP–Kdn. Bioinformatics analysis revealed sequences related to those of the two P. parvum enzymes, suggesting that sialic acid biosynthesis is likely more widespread among microalgae than previously thought and that this acidic sugar may play a role in host–pathogen interactions involving microalgae. Our findings provide evidence that P. parvum has the biosynthetic machinery for de novo production of the deaminated sialic acid Kdn and that sialic acid biosynthesis may be common among microalgae.

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Section: Methodsmentioning

confidence: 99%

Identification of a Kdn biosynthesis pathway in the haptophyte Prymnesium parvum suggests widespread sialic acid biosynthesis among microalgae

Wagstaff

Rejzek

Field

2018

Journal of Biological Chemistry

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“…In 2015, Field et al reported a one-pot multienzyme approach to base modified UDP-Glc and UDP-Gal (Scheme 3). 8 This provided enzymatic access to a new class of 5-position modified sugar-nucleotides, which had previously demonstrated promise as GT inhibitors 9 and as fluorescent tools for GT assay development. 10 Their strategy employed a small panel of 5position modified UTPs and Glc 1-phosphate, incubated with UDP-glucose pyrophosphorylase (GalU) to generate, in situ, a modified UDP-Glc in catalytic quantity, to be continuously recycled by GalU.…”

Section: Towards Udp and Dtdp Sugar-nucleotidesmentioning

confidence: 99%

Recent advances in the enzymatic synthesis of sugar-nucleotides using nucleotidylyltransferases and glycosyltransferases

Ahmadipour

Beswick

Miller

2018

Carbohydrate Research

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“…However,i ng eneral, low yields (5-23 %) were obtained and the conversion of the gluco compounds into the corresponding galacto derivatives was the problematic step. [68] As lightly modified strategy was employed for the preparation of UDPÀGal analogues 80 and 81.I odination of UMP 76 afforded 5-iodo derivative 64,w hichw as transformed into key intermediate 69.M orpholidate 69 was submitted to aS uzuki-Miyaura cross-coupling reactionwith the corresponding boronic acid to furnish compounds 77 and 78,w hich were coupled with d-galactose phosphate to afford compounds 79 and 80, respectively (Scheme12). [69] By comparing the inhibitory activities of compounds 79 and 80 towards a-a nd b-(1,4)GalTsw ith those of 5-substitutedu ridine derivatives 81 and 82 ( Figure 10), which had been prepared from 5-iodouridine 63,i tw as clear that derivatives that incorporated the sugardiphosphate unit improved the results.…”

Section: Modification At the Nucleobasementioning

confidence: 99%

Nucleoside Diphosphate Sugar Analogues that Target Glycosyltransferases

et al. 2016

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Glycosyltransferases (GTs) are enzymes that transfer carbohydrates to ar ange of substrates and they play af undamental role in the recognition processes associated with severald iseases.T he design of glycomimeticso fn ucleoside diphosphate( NDP) sugars-the natural substrates that act as inhibitors or modulators of GTs-is not only necessary for the development of glycan-based therapeutic drugs, but is also ac rucial toolf or understanding their mechanismso fa ction.W hilst al arge number of inhibitors that consist of modifications of the carbohydrate unit and the pyrophosphate linkageh ave been designed,l ess attention has been paidt om odifications at the ribose moiety and the nucleobase. However,i nr ecent years, promising results have been obtainedt hrough such variations, which were prompted by the initial interest in the photolabeling of enzymes to study their structure. In this Focus Review,w e survey recentp rogress in the synthesis of NDP-sugar analogues that are modified at the ribose moiety or at the heterocyclic base.The ORCID identification number(s) for the author(s) of this articlecan be found under http://dx.

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Enzymatic synthesis of nucleobase-modified UDP-sugars: scope and limitations

Abstract: Graphical abstract

Cited by 22 publications

References 28 publications

Identification of a Kdn biosynthesis pathway in the haptophyte Prymnesium parvum suggests widespread sialic acid biosynthesis among microalgae

Identification of a Kdn biosynthesis pathway in the haptophyte Prymnesium parvum suggests widespread sialic acid biosynthesis among microalgae

Recent advances in the enzymatic synthesis of sugar-nucleotides using nucleotidylyltransferases and glycosyltransferases

Nucleoside Diphosphate Sugar Analogues that Target Glycosyltransferases

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