BRENDA - Enzyme Database show
show all sequences of 2.7.8.33

Functional characterization and membrane topology of Escherichia coli WecA, a sugar-phosphate transferase initiating the biosynthesis of enterobacterial common antigen and O-antigen lipopolysaccharide

Lehrer, J.; Vigeant, K.A.; Tatar, L.D.; Valvano, M.A.; J. Bacteriol. 189, 2618-2628 (2007)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
-
Escherichia coli
Engineering
Amino acid exchange
Commentary
Organism
D156E
no detectable transfer activity
Escherichia coli
D156N
no detectable transfer activity
Escherichia coli
D159E
transfer activity is drastically reduced, compared to wild-type enzyme
Escherichia coli
D159N
transfer activity is drastically reduced, compared to wild-type enzyme
Escherichia coli
D90E
slightly reduced velocities and small increases in the apparent Km for UDPGlcNAc. In membranes containing the D90E form of WecA, the apparent Km values for Mg2+ and Mn2+ increases 3-5fold compared to the apparent Km of the parental enzyme
Escherichia coli
D90N
slightly reduced velocities and small increases in the apparent Km for UDPGlcNAc. In membranes containing the D90N form of WecA, the apparent Km values for Mg2+ and Mn2+ increases 3-5fold compared to the apparent Km of the parental enzyme
Escherichia coli
D91E
membranes containing WecA-D91E exhibit a 6fold decrease in the apparent Km for UDP-GlcNAc compared to the apparent Km of the wild-type enzyme. In membranes containing the D91E form of WecA, the apparent Km values with Mg2+ decreases 3fold, compared to the apparent Km of the wild-type parental enzyme
Escherichia coli
D91N
in membranes containing the D91N form of WecA, the apparent Km values with Mg2+ increases 3fold, compared to the apparent Km of the wild-type parental enzyme
Escherichia coli
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.00012
-
UDP-N-acetyl-D-glucosamine
pH 8.5, 37C, in presence of Mg2+
Escherichia coli
0.00019
-
UDP-N-acetyl-D-glucosamine
pH 8.5, 37C, in presence of Mn2+
Escherichia coli
Localization
Localization
Commentary
Organism
GeneOntology No.
Textmining
membrane
integral membrane protein. the C terminus of WecA is exposed to the cytosol. Localizes to discrete regions in the bacterial plasma membrane
Escherichia coli
16020
-
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Mg2+
either Mg2+ or Mn2+ activates the enzyme. In vitro and in the presence of excess UDP-GlcNAc, the enzyme is nearly six times more effective with Mn2+ than with Mg2+. KM-values for wild-type and mutant enzymes
Escherichia coli
Mn2+
either Mg2+ or Mn2+ activates the enzyme. In vitro and in the presence of excess UDP-GlcNAc, the enzyme is nearly six times more effective with Mn2+ than with Mg2+. KM-values for wild-type and mutant enzymes
Escherichia coli
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
38000
-
SDS-PAGE
Escherichia coli
40957
-
x * 40957, calculated from sequence
Escherichia coli
40960
-
calculated from sequence
Escherichia coli
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
UDP-N-acetyl-D-glucosamine + ditrans,polycis-undecaprenyl phosphate
Escherichia coli
initiates the biosynthesis of enterobacterial common antigen and O-antigen lipopolysaccharide
UMP + N-acetyl-D-glucosaminyldiphospho-ditrans,polycis-undecaprenol
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Escherichia coli
P0AC78
-
-
Purification (Commentary)
Commentary
Organism
-
Escherichia coli
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
UDP-N-acetyl-D-glucosamine + ditrans,polycis-undecaprenyl phosphate
initiates the biosynthesis of enterobacterial common antigen and O-antigen lipopolysaccharide
708918
Escherichia coli
UMP + N-acetyl-D-glucosaminyldiphospho-ditrans,polycis-undecaprenol
-
-
-
?
UDP-N-acetyl-D-glucosamine + ditrans,polycis-undecaprenyl phosphate
Asp156 is required for catalysis
708918
Escherichia coli
UMP + N-acetyl-D-glucosaminyldiphospho-ditrans,polycis-undecaprenol
-
-
-
?
Subunits
Subunits
Commentary
Organism
?
x * 38000, SDS-PAGE; x * 40957, calculated from sequence
Escherichia coli
Temperature Optimum [C]
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
37
-
assay at
Escherichia coli
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8.5
-
assay at
Escherichia coli
pI Value
Organism
Commentary
pI Value Maximum
pI Value
Escherichia coli
calculated from sequence
-
10.01
Cloned(Commentary) (protein specific)
Commentary
Organism
-
Escherichia coli
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
D156E
no detectable transfer activity
Escherichia coli
D156N
no detectable transfer activity
Escherichia coli
D159E
transfer activity is drastically reduced, compared to wild-type enzyme
Escherichia coli
D159N
transfer activity is drastically reduced, compared to wild-type enzyme
Escherichia coli
D90E
slightly reduced velocities and small increases in the apparent Km for UDPGlcNAc. In membranes containing the D90E form of WecA, the apparent Km values for Mg2+ and Mn2+ increases 3-5fold compared to the apparent Km of the parental enzyme
Escherichia coli
D90N
slightly reduced velocities and small increases in the apparent Km for UDPGlcNAc. In membranes containing the D90N form of WecA, the apparent Km values for Mg2+ and Mn2+ increases 3-5fold compared to the apparent Km of the parental enzyme
Escherichia coli
D91E
membranes containing WecA-D91E exhibit a 6fold decrease in the apparent Km for UDP-GlcNAc compared to the apparent Km of the wild-type enzyme. In membranes containing the D91E form of WecA, the apparent Km values with Mg2+ decreases 3fold, compared to the apparent Km of the wild-type parental enzyme
Escherichia coli
D91N
in membranes containing the D91N form of WecA, the apparent Km values with Mg2+ increases 3fold, compared to the apparent Km of the wild-type parental enzyme
Escherichia coli
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.00012
-
UDP-N-acetyl-D-glucosamine
pH 8.5, 37C, in presence of Mg2+
Escherichia coli
0.00019
-
UDP-N-acetyl-D-glucosamine
pH 8.5, 37C, in presence of Mn2+
Escherichia coli
Localization (protein specific)
Localization
Commentary
Organism
GeneOntology No.
Textmining
membrane
integral membrane protein. the C terminus of WecA is exposed to the cytosol. Localizes to discrete regions in the bacterial plasma membrane
Escherichia coli
16020
-
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Mg2+
either Mg2+ or Mn2+ activates the enzyme. In vitro and in the presence of excess UDP-GlcNAc, the enzyme is nearly six times more effective with Mn2+ than with Mg2+. KM-values for wild-type and mutant enzymes
Escherichia coli
Mn2+
either Mg2+ or Mn2+ activates the enzyme. In vitro and in the presence of excess UDP-GlcNAc, the enzyme is nearly six times more effective with Mn2+ than with Mg2+. KM-values for wild-type and mutant enzymes
Escherichia coli
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
38000
-
SDS-PAGE
Escherichia coli
40957
-
x * 40957, calculated from sequence
Escherichia coli
40960
-
calculated from sequence
Escherichia coli
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
UDP-N-acetyl-D-glucosamine + ditrans,polycis-undecaprenyl phosphate
Escherichia coli
initiates the biosynthesis of enterobacterial common antigen and O-antigen lipopolysaccharide
UMP + N-acetyl-D-glucosaminyldiphospho-ditrans,polycis-undecaprenol
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
-
Escherichia coli
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
UDP-N-acetyl-D-glucosamine + ditrans,polycis-undecaprenyl phosphate
initiates the biosynthesis of enterobacterial common antigen and O-antigen lipopolysaccharide
708918
Escherichia coli
UMP + N-acetyl-D-glucosaminyldiphospho-ditrans,polycis-undecaprenol
-
-
-
?
UDP-N-acetyl-D-glucosamine + ditrans,polycis-undecaprenyl phosphate
Asp156 is required for catalysis
708918
Escherichia coli
UMP + N-acetyl-D-glucosaminyldiphospho-ditrans,polycis-undecaprenol
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
?
x * 38000, SDS-PAGE; x * 40957, calculated from sequence
Escherichia coli
Temperature Optimum [C] (protein specific)
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
37
-
assay at
Escherichia coli
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8.5
-
assay at
Escherichia coli
pI Value (protein specific)
Organism
Commentary
pI Value Maximum
pI Value
Escherichia coli
calculated from sequence
-
10.01
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
-
-
1
-
1
-
1
-
1
1
-
2
-
2
-
-
1
1
-
-
-
-
6
-
1
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
1
-
-
1
-
-
1
1
-
2
-
-
-
1
-
-
-
-
6
-
1
-
-
-
1
-
-
-
-
3
3
-
-
-
738596
Ishizaki
Inhibition of the first step i ...
Bacillus subtilis 168, Bacillus subtilis
J. Biol. Chem.
288
30309-30319
2013
-
-
-
-
1
-
3
-
-
-
-
2
-
4
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
1
-
2
3
-
-
-
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
723725
Furlong
Characterization of the highly ...
Escherichia coli
Protein Sci.
21
1366-1375
2012
-
-
1
-
9
-
1
2
1
1
-
-
-
2
-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
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-
-
-
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1
-
-
9
-
-
1
-
2
1
1
-
-
-
-
-
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
722351
Hug
Analogies and homologies in li ...
Escherichia coli
Glycobiology
21
138-151
2011
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
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-
-
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-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
710401
Hug
Helicobacter pylori lipopolysa ...
Helicobacter pylori
PLoS Pathog.
6
e1000819
2010
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
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-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
707104
Al-Dabbagh
Preparative enzymatic synthesi ...
Thermotoga maritima
Anal. Biochem.
391
163-165
2009
-
-
-
-
-
-
-
-
1
-
-
1
-
1
-
-
1
-
-
-
-
-
3
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
1
-
-
-
-
3
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
708920
Al-Dabbagh
Purification and characterizat ...
Thermotoga maritima, Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589
J. Bacteriol.
190
7141-7146
2008
2
-
1
-
-
-
4
2
1
1
3
2
-
2
-
-
1
-
-
-
1
-
12
1
1
1
-
2
1
1
-
-
-
-
-
2
-
1
-
-
-
-
-
4
-
2
1
1
3
2
-
-
-
1
-
-
1
-
12
1
1
1
-
2
1
1
-
-
-
-
-
-
-
-
708918
Lehrer
Functional characterization an ...
Escherichia coli
J. Bacteriol.
189
2618-2628
2007
-
-
1
-
8
-
-
2
1
2
3
1
-
1
-
-
1
-
-
-
-
-
2
1
1
-
-
-
1
-
-
-
-
1
-
-
-
1
-
-
8
-
-
-
-
2
1
2
3
1
-
-
-
1
-
-
-
-
2
1
1
-
-
-
1
-
-
1
-
-
-
-
-
-
709837
Amer
Conserved amino acid residues ...
Escherichia coli
Microbiology
147
3015-3025
2001
-
-
1
-
-
-
-
-
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
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-
-
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1
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-
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1
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-
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-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
708621
Rush
Polyisoprenyl phosphate specif ...
Escherichia coli
Glycobiology
7
315-322
1997
-
-
-
-
-
-
1
1
1
1
-
-
-
6
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
1
1
-
-
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
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-
-
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