BRENDA - Enzyme Database show
show all sequences of 2.7.8.33

Catalytic mechanism of MraY and WecA, two paralogues of the polyprenyl-phosphate N-acetylhexosamine 1-phosphate transferase superfamily

Al-Dabbagh, B.; Olatunji, S.; Crouvoisier, M.; El Ghachi, M.; Blanot, D.; Mengin-Lecreulx, D.; Bouhss, A.; Biochimie 127, 249-257 (2016)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
gene wecA, cloning in Escherichia coli strain DH5alpha
Thermotoga maritima
Engineering
Amino acid exchange
Commentary
Organism
D72A
site-directed mutagenesis, very low activity of the mutant protein at pH 8.0, which represents only about 1.2% of the wild-type activity. At pH 7.0, the mutant protein is totally inactive. Increasing the pH from 8 to 9 results in a 2.5fold increase of the D72A mutant activity, while the wild-type enzyme activity rather decreases, from 310 to 240 U/mg of protein
Thermotoga maritima
Inhibitors
Inhibitors
Commentary
Organism
Structure
dodecylamine
-
Thermotoga maritima
Localization
Localization
Commentary
Organism
GeneOntology No.
Textmining
cell wall
-
Thermotoga maritima
5618
-
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Mg2+
essentially required
Thermotoga maritima
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
UDP-N-acetyl-alpha-D-glucosamine + ditrans,octacis-undecaprenyl phosphate
Thermotoga maritima
the enzyme is highly specific for UDP-GlcNAc, its physiological substrate
UMP + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol
-
-
r
UDP-N-acetyl-alpha-D-glucosamine + ditrans,octacis-undecaprenyl phosphate
Thermotoga maritima ATCC 43589
the enzyme is highly specific for UDP-GlcNAc, its physiological substrate
UMP + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol
-
-
r
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Thermotoga maritima
Q9X1N5
gene wecA
-
Thermotoga maritima ATCC 43589
Q9X1N5
gene wecA
-
Purification (Commentary)
Commentary
Organism
recombinant wild-type and D72A mutant enzymes
Thermotoga maritima
Reaction
Reaction
Commentary
Organism
UDP-N-acetyl-alpha-D-glucosamine + ditrans,octacis-undecaprenyl phosphate = UMP + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol
a one-step, single displacement mechanism. The oxyanion of the polyprenyl-phosphate attacks the beta-phosphate of the nucleotide substrate, leading to the formation of lipid product and the liberation of UMP. The involvement of an invariant aspartyl residue in the deprotonation of the lipid substrate is possible
Thermotoga maritima
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
additional information
the enzyme does not display any diphosphatase activity on the nucleotide substrate. Enzyme catalytic mechanism and substrate specificity, overview. The minimal length of the carbon chain of the lipid substrate for an efficient catalysis is 35. The essential aspartate residue, that is invariant in the superfamily, is Asp72 in Thermotoga maritima WecA, the residue is involved in the deprotonation of the lipid substrate during the catalytic process. No activity by the enzyme with UDP-galactose, UDP-GalNAc, GDP-glucose, ADP-ribose, UDP-glucuronic acid, GDP-D-mannose, and UDP-hexanolamine
737788
Thermotoga maritima
?
-
-
-
-
additional information
the enzyme does not display any diphosphatase activity on the nucleotide substrate. Enzyme catalytic mechanism and substrate specificity, overview. The minimal length of the carbon chain of the lipid substrate for an efficient catalysis is 35. The essential aspartate residue, that is invariant in the superfamily, is Asp72 in Thermotoga maritima WecA, the residue is involved in the deprotonation of the lipid substrate during the catalytic process. No activity by the enzyme with UDP-galactose, UDP-GalNAc, GDP-glucose, ADP-ribose, UDP-glucuronic acid, GDP-D-mannose, and UDP-hexanolamine
737788
Thermotoga maritima ATCC 43589
?
-
-
-
-
UDP-N-acetyl-alpha-D-glucosamine + ditrans,octacis-undecaprenyl phosphate
the enzyme is highly specific for UDP-GlcNAc, its physiological substrate
737788
Thermotoga maritima
UMP + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol
-
-
-
r
UDP-N-acetyl-alpha-D-glucosamine + ditrans,octacis-undecaprenyl phosphate
the forward and reverse exchange reactions required the presence of the second substrate, undecaprenyl phosphate and UMP, respectively. The nucleotide substrate UDPMurNAc-pentapeptide, as well as the nucleotide product UMP, can bind to MraY in the absence of lipid ligands. The enzyme is highly specific for UDP-GlcNAc, its physiological substrate
737788
Thermotoga maritima
UMP + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol
-
-
-
r
UDP-N-acetyl-alpha-D-glucosamine + ditrans,octacis-undecaprenyl phosphate
the enzyme is highly specific for UDP-GlcNAc, its physiological substrate
737788
Thermotoga maritima ATCC 43589
UMP + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol
-
-
-
r
UDP-N-acetyl-alpha-D-glucosamine + ditrans,octacis-undecaprenyl phosphate
the forward and reverse exchange reactions required the presence of the second substrate, undecaprenyl phosphate and UMP, respectively. The nucleotide substrate UDPMurNAc-pentapeptide, as well as the nucleotide product UMP, can bind to MraY in the absence of lipid ligands. The enzyme is highly specific for UDP-GlcNAc, its physiological substrate
737788
Thermotoga maritima ATCC 43589
UMP + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol
-
-
-
r
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
65
-
assay at
Thermotoga maritima
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8
-
assay at
Thermotoga maritima
Cloned(Commentary) (protein specific)
Commentary
Organism
gene wecA, cloning in Escherichia coli strain DH5alpha
Thermotoga maritima
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
D72A
site-directed mutagenesis, very low activity of the mutant protein at pH 8.0, which represents only about 1.2% of the wild-type activity. At pH 7.0, the mutant protein is totally inactive. Increasing the pH from 8 to 9 results in a 2.5fold increase of the D72A mutant activity, while the wild-type enzyme activity rather decreases, from 310 to 240 U/mg of protein
Thermotoga maritima
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
dodecylamine
-
Thermotoga maritima
Localization (protein specific)
Localization
Commentary
Organism
GeneOntology No.
Textmining
cell wall
-
Thermotoga maritima
5618
-
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Mg2+
essentially required
Thermotoga maritima
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
UDP-N-acetyl-alpha-D-glucosamine + ditrans,octacis-undecaprenyl phosphate
Thermotoga maritima
the enzyme is highly specific for UDP-GlcNAc, its physiological substrate
UMP + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol
-
-
r
UDP-N-acetyl-alpha-D-glucosamine + ditrans,octacis-undecaprenyl phosphate
Thermotoga maritima ATCC 43589
the enzyme is highly specific for UDP-GlcNAc, its physiological substrate
UMP + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol
-
-
r
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant wild-type and D72A mutant enzymes
Thermotoga maritima
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
additional information
the enzyme does not display any diphosphatase activity on the nucleotide substrate. Enzyme catalytic mechanism and substrate specificity, overview. The minimal length of the carbon chain of the lipid substrate for an efficient catalysis is 35. The essential aspartate residue, that is invariant in the superfamily, is Asp72 in Thermotoga maritima WecA, the residue is involved in the deprotonation of the lipid substrate during the catalytic process. No activity by the enzyme with UDP-galactose, UDP-GalNAc, GDP-glucose, ADP-ribose, UDP-glucuronic acid, GDP-D-mannose, and UDP-hexanolamine
737788
Thermotoga maritima
?
-
-
-
-
additional information
the enzyme does not display any diphosphatase activity on the nucleotide substrate. Enzyme catalytic mechanism and substrate specificity, overview. The minimal length of the carbon chain of the lipid substrate for an efficient catalysis is 35. The essential aspartate residue, that is invariant in the superfamily, is Asp72 in Thermotoga maritima WecA, the residue is involved in the deprotonation of the lipid substrate during the catalytic process. No activity by the enzyme with UDP-galactose, UDP-GalNAc, GDP-glucose, ADP-ribose, UDP-glucuronic acid, GDP-D-mannose, and UDP-hexanolamine
737788
Thermotoga maritima ATCC 43589
?
-
-
-
-
UDP-N-acetyl-alpha-D-glucosamine + ditrans,octacis-undecaprenyl phosphate
the enzyme is highly specific for UDP-GlcNAc, its physiological substrate
737788
Thermotoga maritima
UMP + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol
-
-
-
r
UDP-N-acetyl-alpha-D-glucosamine + ditrans,octacis-undecaprenyl phosphate
the forward and reverse exchange reactions required the presence of the second substrate, undecaprenyl phosphate and UMP, respectively. The nucleotide substrate UDPMurNAc-pentapeptide, as well as the nucleotide product UMP, can bind to MraY in the absence of lipid ligands. The enzyme is highly specific for UDP-GlcNAc, its physiological substrate
737788
Thermotoga maritima
UMP + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol
-
-
-
r
UDP-N-acetyl-alpha-D-glucosamine + ditrans,octacis-undecaprenyl phosphate
the enzyme is highly specific for UDP-GlcNAc, its physiological substrate
737788
Thermotoga maritima ATCC 43589
UMP + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol
-
-
-
r
UDP-N-acetyl-alpha-D-glucosamine + ditrans,octacis-undecaprenyl phosphate
the forward and reverse exchange reactions required the presence of the second substrate, undecaprenyl phosphate and UMP, respectively. The nucleotide substrate UDPMurNAc-pentapeptide, as well as the nucleotide product UMP, can bind to MraY in the absence of lipid ligands. The enzyme is highly specific for UDP-GlcNAc, its physiological substrate
737788
Thermotoga maritima ATCC 43589
UMP + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol
-
-
-
r
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
65
-
assay at
Thermotoga maritima
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8
-
assay at
Thermotoga maritima
General Information
General Information
Commentary
Organism
evolution
the enzyme is a member of the polyprenyl-phosphate N-acetylhexosamine 1-phosphate transferase, P2HPT, superfamily
Thermotoga maritima
metabolism
WecA catalyzes the first membrane step of the biosynthesis of many cell wall polymers such as the O-antigen, teichoic acids and arabinogalactan
Thermotoga maritima
physiological function
enzyme WecA, catalyzes the transfer of the phospho-GlcNAc moiety of UDP-N-acetylglucosamine onto the same lipid carrier, leading to the formation of N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol that is essential for the synthesis of various bacterial cell envelope components. Undecaprenyl diphosphoryl-N-acetylglucosamine (lipid intermediate I) is a ubiquitous compound in all kingdoms of life. the enzyme is involved in the initiation of enterobacterial common antigen biosynthesis
Thermotoga maritima
General Information (protein specific)
General Information
Commentary
Organism
evolution
the enzyme is a member of the polyprenyl-phosphate N-acetylhexosamine 1-phosphate transferase, P2HPT, superfamily
Thermotoga maritima
metabolism
WecA catalyzes the first membrane step of the biosynthesis of many cell wall polymers such as the O-antigen, teichoic acids and arabinogalactan
Thermotoga maritima
physiological function
enzyme WecA, catalyzes the transfer of the phospho-GlcNAc moiety of UDP-N-acetylglucosamine onto the same lipid carrier, leading to the formation of N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol that is essential for the synthesis of various bacterial cell envelope components. Undecaprenyl diphosphoryl-N-acetylglucosamine (lipid intermediate I) is a ubiquitous compound in all kingdoms of life. the enzyme is involved in the initiation of enterobacterial common antigen biosynthesis
Thermotoga maritima
Other publictions for EC 2.7.8.33
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
737788
Al-Dabbagh
Catalytic mechanism of MraY an ...
Thermotoga maritima, Thermotoga maritima ATCC 43589
Biochimie
127
249-257
2016
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1
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1
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1
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1
1
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2
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2
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1
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6
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1
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1
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1
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6
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1
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1
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3
3
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738596
Ishizaki
Inhibition of the first step i ...
Bacillus subtilis 168, Bacillus subtilis
J. Biol. Chem.
288
30309-30319
2013
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1
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3
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2
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4
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2
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2
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1
1
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723725
Furlong
Characterization of the highly ...
Escherichia coli
Protein Sci.
21
1366-1375
2012
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1
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9
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1
2
1
1
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2
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1
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1
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9
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1
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1
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1
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722351
Hug
Analogies and homologies in li ...
Escherichia coli
Glycobiology
21
138-151
2011
-
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1
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1
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1
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710401
Hug
Helicobacter pylori lipopolysa ...
Helicobacter pylori
PLoS Pathog.
6
e1000819
2010
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1
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1
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707104
Al-Dabbagh
Preparative enzymatic synthesi ...
Thermotoga maritima
Anal. Biochem.
391
163-165
2009
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1
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3
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1
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1
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708920
Al-Dabbagh
Purification and characterizat ...
Thermotoga maritima, Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589
J. Bacteriol.
190
7141-7146
2008
2
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1
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4
2
1
1
3
2
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2
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1
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1
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12
1
1
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1
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1
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1
1
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708918
Lehrer
Functional characterization an ...
Escherichia coli
J. Bacteriol.
189
2618-2628
2007
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1
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8
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2
1
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3
1
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1
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1
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8
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1
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1
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1
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2
1
1
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1
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1
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709837
Amer
Conserved amino acid residues ...
Escherichia coli
Microbiology
147
3015-3025
2001
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1
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708621
Rush
Polyisoprenyl phosphate specif ...
Escherichia coli
Glycobiology
7
315-322
1997
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1
1
1
1
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6
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3
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