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PMID:18263721

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Citation

Namboori, SC and Graham, DE (2008) Acetamido sugar biosynthesis in the Euryarchaea. J. Bacteriol. 190:2987-96

Abstract

Archaea and eukaryotes share a dolichol phosphate-dependent system for protein N-glycosylation. In both domains, the acetamido sugar N-acetylglucosamine (GlcNAc) forms part of the core oligosaccharide. However, the archaeal Methanococcales produce GlcNAc using the bacterial biosynthetic pathway. Key enzymes in this pathway belong to large families of proteins with diverse functions; therefore, the archaeal enzymes could not be identified solely using comparative sequence analysis. Genes encoding acetamido sugar-biosynthetic proteins were identified in Methanococcus maripaludis using phylogenetic and gene cluster analyses. Proteins expressed in Escherichia coli were purified and assayed for the predicted activities. The MMP1680 protein encodes a universally conserved glucosamine-6-phosphate synthase. The MMP1077 phosphomutase converted alpha-D-glucosamine-6-phosphate to alpha-D-glucosamine-1-phosphate, although this protein is more closely related to archaeal pentose and glucose phosphomutases than to bacterial glucosamine phosphomutases. The thermostable MJ1101 protein catalyzed both the acetylation of glucosamine-1-phosphate and the uridylyltransferase reaction with UTP to produce UDP-GlcNAc. The MMP0705 protein catalyzed the C-2 epimerization of UDP-GlcNAc, and the MMP0706 protein used NAD(+) to oxidize UDP-N-acetylmannosamine, forming UDP-N-acetylmannosaminuronate (ManNAcA). These two proteins are similar to enzymes used for proteobacterial lipopolysaccharide biosynthesis and gram-positive bacterial capsule production, suggesting a common evolutionary origin and a widespread distribution of ManNAcA. UDP-GlcNAc and UDP-ManNAcA biosynthesis evolved early in the euryarchaeal lineage, because most of their genomes contain orthologs of the five genes characterized here. These UDP-acetamido sugars are predicted to be precursors for flagellin and S-layer protein modifications and for the biosynthesis of methanogenic coenzyme B.

Links

PubMed PMC2293230 Online version:10.1128/JB.01970-07

Keywords

Acetylglucosamine/analogs & derivatives; Acetylglucosamine/biosynthesis; Acetylglucosamine/metabolism; Archaeal Proteins/genetics; Archaeal Proteins/isolation & purification; Archaeal Proteins/metabolism; Biosynthetic Pathways; Carbohydrate Dehydrogenases/metabolism; Carbohydrate Epimerases/metabolism; Cloning, Molecular; DNA, Archaeal/chemistry; DNA, Archaeal/genetics; Enzymes/genetics; Enzymes/isolation & purification; Enzymes/metabolism; Escherichia coli/genetics; Glucosamine/analogs & derivatives; Glucosamine/metabolism; Glucose-6-Phosphate/analogs & derivatives; Glucose-6-Phosphate/metabolism; Glucosephosphates/metabolism; Glutamine-Fructose-6-Phosphate Transaminase (Isomerizing)/genetics; Glutamine-Fructose-6-Phosphate Transaminase (Isomerizing)/metabolism; Methanococcus/genetics; Methanococcus/metabolism; Molecular Sequence Data; NAD/metabolism; Phosphotransferases (Phosphomutases)/genetics; Phosphotransferases (Phosphomutases)/metabolism; Sequence Analysis, DNA; Uridine Diphosphate N-Acetylglucosamine/metabolism; Uridine Diphosphate Sugars/metabolism; Uridine Triphosphate/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

METMP:GLMS

enables

GO:0004360: glutamine-fructose-6-phosphate transaminase (isomerizing) activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

METMP:GLMS

GO:0004360: glutamine-fructose-6-phosphate transaminase (isomerizing) activity

ECO:0000314:

F

Fig 4: formation of Glucoosamine-6-phosphate

complete
CACAO 6652

METMP:WECB

enables

GO:0008761: UDP-N-acetylglucosamine 2-epimerase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

METMP:WECB

GO:0008761: UDP-N-acetylglucosamine 2-epimerase activity

ECO:0000314:

F

Fig 6: The MMP0705 protein has epimerase activity

complete
CACAO 6656

METMP:GLMM

enables

GO:0008966: phosphoglucosamine mutase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

METMP:GLMM

GO:0008966: phosphoglucosamine mutase activity

ECO:0000314:

F

Fig 5A: when MMP1077 is involved in the reaction UDP-GlcNAc is produced

complete
CACAO 6664

METJA:GLMU

enables

GO:0003977: UDP-N-acetylglucosamine diphosphorylase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

METJA:GLMU

enables

GO:0019134: glucosamine-1-phosphate N-acetyltransferase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

METMP:WECC

GO:0016616 : oxidoreductase activity, acting on the CH-OH group of donors, NAD or NADP as acceptor

ECO:0000314:

F

figure 6:oxidizes UDP-ManNAc into UDP-ManNAcA by using NAD+ resulting in 2 NADH

complete
CACAO 6673

METMP:WECC

enables

GO:0016616: oxidoreductase activity, acting on the CH-OH group of donors, NAD or NADP as acceptor

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete


See also

References

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