PMID- 28096310
OWN - NLM
STAT- MEDLINE
DCOM- 20180102
LR  - 20181113
IS  - 1460-2423 (Electronic)
IS  - 0959-6658 (Print)
IS  - 0959-6658 (Linking)
VI  - 27
IP  - 4
DP  - 2017 Apr 1
TI  - Characterization of the dTDP-Fuc3N and dTDP-Qui3N biosynthetic pathways in 
      Campylobacter jejuni 81116.
PG  - 358-369
LID - 10.1093/glycob/cww136 [doi]
AB  - The Gram-negative bacterium Campylobacter jejuni 81116 (Penner serotype HS:6) has 
      a class E lipooligosaccharide (LOS) biosynthesis locus containing 19 genes, which 
      encode for 11 putative glycosyltransferases, 1 lipid A acyltransferase and 7 
      enzymes thought to be involved in the biosynthesis of dideoxyhexosamine (ddHexN) 
      moieties. Although the LOS outer core structure of C. jejuni 81116 is still 
      unknown, recent mass spectrometry analyses suggest that it contains acetylated 
      forms of two ddHexN residues. For this investigation, five of the genes encoding 
      enzymes reportedly involved in the biosyntheses of these sugar residues were 
      examined, rmlA, rmlB, wlaRA, wlaRB and wlaRG. Specifically, these genes were 
      cloned and expressed in Escherichia coli, and the corresponding enzymes were 
      purified and tested for biochemical activity. Here we present data demonstrating 
      that RmlA functions as a glucose-1-phosphate thymidylyltransferase and that RmlB 
      is a thymidine diphosphate (dTDP)-glucose 4,6-dehydratase. We also show, through 
      nuclear magnetic resonance spectroscopy and mass spectrometry analyses, that 
      WlaRG, when utilized in coupled assays with either WlaRA or WlaRB and 
      dTDP-4-keto-6-deoxyglucose, results in the production of either 
      dTDP-3-amino-3,6-dideoxy-d-galactose (dTDP-Fuc3N) or 
      dTDP-3-amino-3,6-dideoxy-d-glucose (dTDP-Qui3N), respectively. In addition, the 
      X-ray crystallographic structures of the 3,4-ketoisomerases, WlaRA and WlaRB, 
      were determined to 2.14 and 2.0 A resolutions, respectively. Taken together, the 
      data reported herein demonstrate that C. jejuni 81116 utilizes five enzymes to 
      synthesize dTDP-Fuc3N or dTDP-Qui3N and that WlaRG, an aminotransferase, can 
      function on sugars with differing stereochemistry about their C-4' carbons. 
      Importantly, the data reveal that C. jejuni 81116 has the ability to synthesize 
      two isomeric ddHexN forms.
CI  - (c) Her Majesty the Queen in Right of Canada 2017. Reproduced with the permission 
      of the Minister of National Research Council of Canada.
FAU - Li, Zack Z
AU  - Li ZZ
AD  - National Research Council Canada, Human Health Therapeutics, 100 Sussex Drive, 
      Ottawa, ON, Canada.
FAU - Riegert, Alexander S
AU  - Riegert AS
AD  - Department of Biochemistry, University of Wisconsin, 440 Henry Mall, Madison, WI, 
      USA.
FAU - Goneau, Marie-France
AU  - Goneau MF
AD  - National Research Council Canada, Human Health Therapeutics, 100 Sussex Drive, 
      Ottawa, ON, Canada.
FAU - Cunningham, Anna M
AU  - Cunningham AM
AD  - National Research Council Canada, Human Health Therapeutics, 100 Sussex Drive, 
      Ottawa, ON, Canada.
FAU - Vinogradov, Evgeny
AU  - Vinogradov E
AD  - National Research Council Canada, Human Health Therapeutics, 100 Sussex Drive, 
      Ottawa, ON, Canada.
FAU - Li, Jianjun
AU  - Li J
AD  - National Research Council Canada, Human Health Therapeutics, 100 Sussex Drive, 
      Ottawa, ON, Canada.
FAU - Schoenhofen, Ian C
AU  - Schoenhofen IC
AD  - National Research Council Canada, Human Health Therapeutics, 100 Sussex Drive, 
      Ottawa, ON, Canada.
FAU - Thoden, James B
AU  - Thoden JB
AD  - Department of Biochemistry, University of Wisconsin, 440 Henry Mall, Madison, WI, 
      USA.
FAU - Holden, Hazel M
AU  - Holden HM
AD  - Department of Biochemistry, University of Wisconsin, 440 Henry Mall, Madison, WI, 
      USA.
FAU - Gilbert, Michel
AU  - Gilbert M
AD  - National Research Council Canada, Human Health Therapeutics, 100 Sussex Drive, 
      Ottawa, ON, Canada.
LA  - eng
GR  - R01 GM115921/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PL  - England
TA  - Glycobiology
JT  - Glycobiology
JID - 9104124
RN  - 0 (Lipopolysaccharides)
RN  - 0 (Thymine Nucleotides)
RN  - 0 (lipid-linked oligosaccharides)
RN  - EC 2.3.- (Acyltransferases)
RN  - EC 2.4.- (Glycosyltransferases)
RN  - EC 2.7.7.- (Nucleotidyltransferases)
RN  - EC 2.7.7.24 (glucose-1-phosphate thymidylyltransferase)
RN  - IY9XDZ35W2 (Glucose)
RN  - X2RN3Q8DNE (Galactose)
SB  - IM
MH  - Acyltransferases/chemistry/*genetics/metabolism
MH  - Biosynthetic Pathways/genetics
MH  - Campylobacter jejuni/enzymology/*genetics
MH  - Crystallography, X-Ray
MH  - Escherichia coli/genetics
MH  - Galactose/chemistry/*genetics/metabolism
MH  - Glucose/chemistry/metabolism
MH  - Glycosyltransferases/chemistry/*genetics/metabolism
MH  - Lipopolysaccharides/biosynthesis/genetics
MH  - Nucleotidyltransferases/chemistry/*genetics/metabolism
MH  - Thymine Nucleotides/chemistry/metabolism
PMC - PMC5441919
OTO - NOTNLM
OT  - Campylobacter
OT  - fucosamine
OT  - ketoisomerase
OT  - lipooligosaccharide
OT  - quinovosamine
EDAT- 2017/01/18 06:00
MHDA- 2018/01/03 06:00
PMCR- 2018/04/01
CRDT- 2017/01/19 06:00
PHST- 2016/11/02 00:00 [received]
PHST- 2017/01/11 00:00 [accepted]
PHST- 2017/01/18 06:00 [pubmed]
PHST- 2018/01/03 06:00 [medline]
PHST- 2017/01/19 06:00 [entrez]
PHST- 2018/04/01 00:00 [pmc-release]
AID - cww136 [pii]
AID - 10.1093/glycob/cww136 [doi]
PST - ppublish
SO  - Glycobiology. 2017 Apr 1;27(4):358-369. doi: 10.1093/glycob/cww136.