BRENDA - Enzyme Database
show all sequences of 2.4.99.B6

Structure of the Escherichia coli heptosyltransferase WaaC: binary complexes with ADP and ADP-2-deoxy-2-fluoro heptose

Grizot, S.; Salem, M.; Vongsouthi, V.; Durand, L.; Moreau, F.; Dohi, H.; Vincent, S.; Escaich, S.; Ducruix, A.; J. Mol. Biol. 363, 383-394 (2006)

Data extracted from this reference:

Application
Application
Commentary
Organism
medicine
absence of the enzyme results in a truncated lipopolysaccharide associated with the deeprough phenotype causing a greater susceptibility to antibiotic and an attenuated virulence for pathogenic Gram-negative bacteria. Thus, the enzyme represents a promising target in antibacterial drug design
Escherichia coli
Crystallization (Commentary)
Crystallization (Commentary)
Organism
crystals of the enzyme are grown at 18C by the vapor-diffusion, hanging-drop method by mixing equal volumes of protein (4 mg/ml) and reservoir solution (100 mM Hepes (pH 7.0), 15% (w/v) PEG 1500, 100 mM NaCl (and 5 mM DTT in the case of the Se-Met protein)). Crystals grow within a few days as thin plates (0.15 mM x 0.15 mM x 0.050 mM) and belong to the orthorhombic space group P2(1)2(1)2(1) (a = 78 A, b = 88 A, c = 89 A) with two molecules in the asymmetric unit. Determination of the structure of the enzyme alone at 1.9 A resolution, and in complex with either ADP or the non-cleavable analog adenosine 5'-diphospho-2-deoxy-2-fluoro-L-glycero-beta-D-gluco-heptopyranoside (ADP-2-deoxy-2-fluoro-heptose) of the sugar donor at 2.4 A resolution. Both binary complexes offer a close view of the donor subsite and, together with results from site-directed mutagenesis studies, provide evidence for a model of the catalytic mechanism
Escherichia coli
Inhibitors
Inhibitors
Commentary
Organism
Structure
adenosine 5'-diphospho-2-deoxy-2-fluoro-L-glycero-beta-D-gluco-heptopyranoside
competitive inhibitor
Escherichia coli
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
ADP-L-glycero-beta-D-manno-heptose + (3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->4)-(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->6)-2-deoxy-2-[[(3R)-3-(dodecanoyloxy)tetradecanoyl]amino]-3-O-[(3R)-3-(tetradecanoyloxy)tetradecanoyl]-4-O-phospho-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-O-phosphono-alpha-D-glucopyranose
Escherichia coli
the enzyme is involved in the synthesis of the inner core region of lipopolysaccharide. It catalyzes the addition of the first L-glycero-D-manno-heptose (heptose) molecule to one 3-deoxy-D-manno-oct-2-ulosonic acid residue of the Kdo2-lipid A molecule. Heptose is an essential component of the lipopolysaccharide core domain. Its absence results in a truncated lipopolysaccharide associated with the deeprough phenotype causing a greater susceptibility to antibiotic and an attenuated virulence for pathogenic Gram-negative bacteria
ADP + alpha-L-glycero-D-manno-heptosyl-(1->5)-[(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->4)]-(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->6)-2-deoxy-2-[[(3R)-3-(dodecanoyloxy)tetradecanoyl]amino]-3-O-[(3R)-3-(tetradecanoyloxy)tetradecanoyl]-4-O-phospho-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-O-phosphono-alpha-D-glucopyranose
-
-
?
Organism
Organism
UniProt
Commentary
Textmining
Escherichia coli
P24173
pathogenic strain RS218
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
ADP-L-glycero-beta-D-manno-heptose + (3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->4)-(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->6)-2-deoxy-2-[[(3R)-3-(dodecanoyloxy)tetradecanoyl]amino]-3-O-[(3R)-3-(tetradecanoyloxy)tetradecanoyl]-4-O-phospho-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-O-phosphono-alpha-D-glucopyranose
the enzyme is involved in the synthesis of the inner core region of lipopolysaccharide. It catalyzes the addition of the first L-glycero-D-manno-heptose (heptose) molecule to one 3-deoxy-D-manno-oct-2-ulosonic acid residue of the Kdo2-lipid A molecule. Heptose is an essential component of the lipopolysaccharide core domain. Its absence results in a truncated lipopolysaccharide associated with the deeprough phenotype causing a greater susceptibility to antibiotic and an attenuated virulence for pathogenic Gram-negative bacteria
728139
Escherichia coli
ADP + alpha-L-glycero-D-manno-heptosyl-(1->5)-[(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->4)]-(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->6)-2-deoxy-2-[[(3R)-3-(dodecanoyloxy)tetradecanoyl]amino]-3-O-[(3R)-3-(tetradecanoyloxy)tetradecanoyl]-4-O-phospho-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-O-phosphono-alpha-D-glucopyranose
-
-
-
?
Subunits
Subunits
Commentary
Organism
monomer
-
Escherichia coli
Synonyms
Synonyms
Commentary
Organism
waaC
-
Escherichia coli
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Escherichia coli
IC50 Value
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
0.03
-
pH 7.5, temperature not specified in the publication
Escherichia coli
adenosine 5'-diphospho-2-deoxy-2-fluoro-L-glycero-beta-D-gluco-heptopyranoside
Application (protein specific)
Application
Commentary
Organism
medicine
absence of the enzyme results in a truncated lipopolysaccharide associated with the deeprough phenotype causing a greater susceptibility to antibiotic and an attenuated virulence for pathogenic Gram-negative bacteria. Thus, the enzyme represents a promising target in antibacterial drug design
Escherichia coli
Crystallization (Commentary) (protein specific)
Crystallization
Organism
crystals of the enzyme are grown at 18C by the vapor-diffusion, hanging-drop method by mixing equal volumes of protein (4 mg/ml) and reservoir solution (100 mM Hepes (pH 7.0), 15% (w/v) PEG 1500, 100 mM NaCl (and 5 mM DTT in the case of the Se-Met protein)). Crystals grow within a few days as thin plates (0.15 mM x 0.15 mM x 0.050 mM) and belong to the orthorhombic space group P2(1)2(1)2(1) (a = 78 A, b = 88 A, c = 89 A) with two molecules in the asymmetric unit. Determination of the structure of the enzyme alone at 1.9 A resolution, and in complex with either ADP or the non-cleavable analog adenosine 5'-diphospho-2-deoxy-2-fluoro-L-glycero-beta-D-gluco-heptopyranoside (ADP-2-deoxy-2-fluoro-heptose) of the sugar donor at 2.4 A resolution. Both binary complexes offer a close view of the donor subsite and, together with results from site-directed mutagenesis studies, provide evidence for a model of the catalytic mechanism
Escherichia coli
IC50 Value (protein specific)
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
0.03
-
pH 7.5, temperature not specified in the publication
Escherichia coli
adenosine 5'-diphospho-2-deoxy-2-fluoro-L-glycero-beta-D-gluco-heptopyranoside
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
adenosine 5'-diphospho-2-deoxy-2-fluoro-L-glycero-beta-D-gluco-heptopyranoside
competitive inhibitor
Escherichia coli
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
ADP-L-glycero-beta-D-manno-heptose + (3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->4)-(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->6)-2-deoxy-2-[[(3R)-3-(dodecanoyloxy)tetradecanoyl]amino]-3-O-[(3R)-3-(tetradecanoyloxy)tetradecanoyl]-4-O-phospho-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-O-phosphono-alpha-D-glucopyranose
Escherichia coli
the enzyme is involved in the synthesis of the inner core region of lipopolysaccharide. It catalyzes the addition of the first L-glycero-D-manno-heptose (heptose) molecule to one 3-deoxy-D-manno-oct-2-ulosonic acid residue of the Kdo2-lipid A molecule. Heptose is an essential component of the lipopolysaccharide core domain. Its absence results in a truncated lipopolysaccharide associated with the deeprough phenotype causing a greater susceptibility to antibiotic and an attenuated virulence for pathogenic Gram-negative bacteria
ADP + alpha-L-glycero-D-manno-heptosyl-(1->5)-[(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->4)]-(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->6)-2-deoxy-2-[[(3R)-3-(dodecanoyloxy)tetradecanoyl]amino]-3-O-[(3R)-3-(tetradecanoyloxy)tetradecanoyl]-4-O-phospho-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-O-phosphono-alpha-D-glucopyranose
-
-
?
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
ADP-L-glycero-beta-D-manno-heptose + (3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->4)-(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->6)-2-deoxy-2-[[(3R)-3-(dodecanoyloxy)tetradecanoyl]amino]-3-O-[(3R)-3-(tetradecanoyloxy)tetradecanoyl]-4-O-phospho-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-O-phosphono-alpha-D-glucopyranose
the enzyme is involved in the synthesis of the inner core region of lipopolysaccharide. It catalyzes the addition of the first L-glycero-D-manno-heptose (heptose) molecule to one 3-deoxy-D-manno-oct-2-ulosonic acid residue of the Kdo2-lipid A molecule. Heptose is an essential component of the lipopolysaccharide core domain. Its absence results in a truncated lipopolysaccharide associated with the deeprough phenotype causing a greater susceptibility to antibiotic and an attenuated virulence for pathogenic Gram-negative bacteria
728139
Escherichia coli
ADP + alpha-L-glycero-D-manno-heptosyl-(1->5)-[(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->4)]-(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->6)-2-deoxy-2-[[(3R)-3-(dodecanoyloxy)tetradecanoyl]amino]-3-O-[(3R)-3-(tetradecanoyloxy)tetradecanoyl]-4-O-phospho-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-O-phosphono-alpha-D-glucopyranose
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
monomer
-
Escherichia coli
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Escherichia coli
General Information
General Information
Commentary
Organism
physiological function
the enzyme is involved in the synthesis of the inner core region of lipopolysaccharide. It catalyzes the addition of the first L-glycero-D-manno-heptose (heptose) molecule to one 3-deoxy-D-manno-oct-2-ulosonic acid residue of the Kdo2-lipid A molecule. Heptose is an essential component of the lipopolysaccharide core domain. Its absence results in a truncated lipopolysaccharide associated with the deeprough phenotype causing a greater susceptibility to antibiotic and an attenuated virulence for pathogenic Gram-negative bacteria
Escherichia coli
General Information (protein specific)
General Information
Commentary
Organism
physiological function
the enzyme is involved in the synthesis of the inner core region of lipopolysaccharide. It catalyzes the addition of the first L-glycero-D-manno-heptose (heptose) molecule to one 3-deoxy-D-manno-oct-2-ulosonic acid residue of the Kdo2-lipid A molecule. Heptose is an essential component of the lipopolysaccharide core domain. Its absence results in a truncated lipopolysaccharide associated with the deeprough phenotype causing a greater susceptibility to antibiotic and an attenuated virulence for pathogenic Gram-negative bacteria
Escherichia coli
Other publictions for EC 2.4.99.B6
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
Synonyms
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)
737682
Czyzyk
Escherichia coli heptosyltrans ...
Escherichia coli
Biochemistry
52
5158-5160
2013
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726955
Czyzyk
Lipopolysaccharide biosynthesi ...
Escherichia coli
Biochemistry
50
10570-10572
2011
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8
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1
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2
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6
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1
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8
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8
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1
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6
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8
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1
1
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8
8
727326
Durka
The inhibition of liposacchari ...
Escherichia coli
Chemistry
18
641-651
2011
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1
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1
1
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727135
Moreau
Discovery of new Gram-negative ...
Escherichia coli
Bioorg. Med. Chem. Lett.
18
4022-4026
2008
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1
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11
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1
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1
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1
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10
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1
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11
10
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1
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728139
Grizot
Structure of the Escherichia c ...
Escherichia coli
J. Mol. Biol.
363
383-394
2006
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1
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1
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1
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1
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1
1
1
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1
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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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1
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1
1
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728049
Gronow
Characterization of the physio ...
Escherichia coli
J. Endotoxin Res.
7
263-270
2001
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3
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727413
Gronow
Comparative functional charact ...
Escherichia coli
Eur. J. Biochem.
267
6602-6611
2000
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3
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1
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727824
Kadrmas
Enzymatic synthesis of lipopol ...
Escherichia coli
J. Biol. Chem.
273
2799-2807
1998
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