Structure and mechanism of a bacterial beta -glucosaminidase having O-GlcNAcase activity
Dennis, R.J.; Taylor, E.J.; Macauley, M.S.; Stubbs, K.A.; Turkenburg, J.P.; Hart, S.J.; Black, G.N.; Vocadlo, D.J.; Davies, G.J.; Nat. Struct. Mol. Biol. 13, 365-371 (2006) 
Data extracted from this reference:
Cloned(Commentary)
EC Number
Cloned (Commentary)
Organism
3.2.1.169
soluble BtGH84 is expressed in the cytoplasm of Escherichia coli
Bacteroides thetaiotaomicron
Crystallization (Commentary)
EC Number
Crystallization (Commentary)
Organism
3.2.1.169
X-ray crystal structure of BtGH84 in complex with the inhibitor NAG-thiazoline. Hanging drop vapor diffusion method
Bacteroides thetaiotaomicron
Engineering
EC Number
Protein Variants
Commentary
Organism
3.2.1.169
D242A
site-directed mutant, 2800fold lower catalytic efficiency than wild-type
Bacteroides thetaiotaomicron
3.2.1.169
D242N
site-directed mutant
Bacteroides thetaiotaomicron
3.2.1.169
D243A
site-directed mutant, 110fold lower catalytic efficiency than wild-type
Bacteroides thetaiotaomicron
3.2.1.169
D243N
site-directed mutant
Bacteroides thetaiotaomicron
3.2.1.169
N372A
site-directed mutant
Bacteroides thetaiotaomicron
3.2.1.169
Y137F
site-directed mutant
Bacteroides thetaiotaomicron
3.2.1.169
Y282F
site-directed mutant
Bacteroides thetaiotaomicron
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
3.2.1.169
1,2-dideoxy-2'-methyl-alpha-D-glucopyranoso-[2,1-d]-DELTA2'-thiazoline
NAG-thiazoline
Bacteroides thetaiotaomicron
3.2.1.169
O-(2-acetamido-2-deoxy-D-glucopyranosylidene)amino-N-phenylcarbamate
PUGNAc
Bacteroides thetaiotaomicron
3.2.1.169
streptozotocin
STZ
Bacteroides thetaiotaomicron
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.2.1.169
0.016
-
4-nitrophenyl N-acetyl-beta-D-glucosaminide
D243N mutant
Bacteroides thetaiotaomicron
3.2.1.169
0.092
-
4-nitrophenyl N-acetyl-beta-D-glucosaminide
D243A mutant
Bacteroides thetaiotaomicron
3.2.1.169
0.28
-
4-nitrophenyl N-acetyl-beta-D-glucosaminide
wild-type
Bacteroides thetaiotaomicron
3.2.1.169
0.3
-
4-nitrophenyl N-acetyl-beta-D-glucosaminide
-
Bacteroides thetaiotaomicron
3.2.1.169
0.41
-
4-nitrophenyl N-acetyl-beta-D-glucosaminide
Y282F mutant
Bacteroides thetaiotaomicron
3.2.1.169
1.18
-
4-nitrophenyl N-acetyl-beta-D-glucosaminide
Y137F mutant
Bacteroides thetaiotaomicron
3.2.1.169
1.4
-
4-nitrophenyl N-acetyl-beta-D-thioglucosaminide
-
Bacteroides thetaiotaomicron
3.2.1.169
1.47
-
4-nitrophenyl N-acetyl-beta-D-glucosaminide
D242N mutant
Bacteroides thetaiotaomicron
3.2.1.169
4.4
-
4-nitrophenyl N-acetyl-beta-D-glucosaminide
D242A mutant
Bacteroides thetaiotaomicron
3.2.1.169
36
-
4-nitrophenyl N-acetyl-beta-D-glucosaminide
N372A mutant
Bacteroides thetaiotaomicron
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
3.2.1.169
additional information
Bacteroides thetaiotaomicron
enzyme cleaves 4-nitrophenyl N-acetyl-beta-D-glucosaminide from modified proteins and also cleaves 4-nitrophenyl N-acetyl-beta-D-thioglucosaminide
?
-
-
?
Organism
EC Number
Organism
UniProt
Commentary
Textmining
3.2.1.147
Bacteroides thetaiotaomicron
-
-
-
3.2.1.169
Bacteroides thetaiotaomicron
Q89ZI2
-
-
Purification (Commentary)
EC Number
Purification (Commentary)
Organism
3.2.1.169
affinity chromatography using nickel-affinity chromatography followed by gel filtration
Bacteroides thetaiotaomicron
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
3.2.1.147
4-nitrophenyl 2-acetamido-2-deoxy-1-thio-beta-D-glucopyranoside + H2O
-
705869
Bacteroides thetaiotaomicron
4-nitrobenzenethiol + N-acetyl-D-glucosamine
-
-
-
?
3.2.1.169
4-methylumbelliferyl N-acetyl-beta-D-glucosaminide + H2O
-
705869
Bacteroides thetaiotaomicron
4-methylumbelliferone + N-acetyl-beta-D-glucosamine
-
-
-
?
3.2.1.169
4-methylumbelliferyl N-difluoroacetyl-beta-D-glucosaminide + H2O
-
705869
Bacteroides thetaiotaomicron
4-methylumbelliferone + N-difluoroacetyl-beta-D-glucosamine
-
-
-
?
3.2.1.169
4-methylumbelliferyl N-fluoroacetyl-beta-D-glucosaminide + H2O
-
705869
Bacteroides thetaiotaomicron
4-methylumbelliferone + N-fluoroacetyl-beta-D-glucosamine
-
-
-
?
3.2.1.169
4-methylumbelliferyl N-trifluoroacetyl-beta-D-glucosaminide + H2O
-
705869
Bacteroides thetaiotaomicron
4-methylumbelliferone + N-trifluoroacetyl-beta-D-glucosamine
-
-
-
?
3.2.1.169
4-nitrophenyl N-acetyl-beta-D-glucosaminide + H2O
-
705869
Bacteroides thetaiotaomicron
4-nitrophenol + N-acetyl-beta-D-glucosamine
-
-
-
?
3.2.1.169
4-nitrophenyl N-acetyl-beta-D-thioglucosaminide + H2O
-
705869
Bacteroides thetaiotaomicron
4-nitrophenol + N-acetyl-beta-D-thioglucosamine
-
-
-
?
3.2.1.169
additional information
enzyme cleaves 4-nitrophenyl N-acetyl-beta-D-glucosaminide from modified proteins and also cleaves 4-nitrophenyl N-acetyl-beta-D-thioglucosaminide
705869
Bacteroides thetaiotaomicron
?
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
3.2.1.169
dimer
-
Bacteroides thetaiotaomicron
Synonyms
EC Number
Synonyms
Commentary
Organism
3.2.1.147
thioglycosidase
-
Bacteroides thetaiotaomicron
3.2.1.169
BtGH84
-
Bacteroides thetaiotaomicron
3.2.1.169
O-GlcNAcase
-
Bacteroides thetaiotaomicron
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
3.2.1.169
37
-
assay at
Bacteroides thetaiotaomicron
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.2.1.147
7.4
-
assay at
Bacteroides thetaiotaomicron
3.2.1.169
6
-
bell-shaped pH-activity profile
Bacteroides thetaiotaomicron
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
3.2.1.169
soluble BtGH84 is expressed in the cytoplasm of Escherichia coli
Bacteroides thetaiotaomicron
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
3.2.1.169
X-ray crystal structure of BtGH84 in complex with the inhibitor NAG-thiazoline. Hanging drop vapor diffusion method
Bacteroides thetaiotaomicron
Engineering (protein specific)
EC Number
Protein Variants
Commentary
Organism
3.2.1.169
D242A
site-directed mutant, 2800fold lower catalytic efficiency than wild-type
Bacteroides thetaiotaomicron
3.2.1.169
D242N
site-directed mutant
Bacteroides thetaiotaomicron
3.2.1.169
D243A
site-directed mutant, 110fold lower catalytic efficiency than wild-type
Bacteroides thetaiotaomicron
3.2.1.169
D243N
site-directed mutant
Bacteroides thetaiotaomicron
3.2.1.169
N372A
site-directed mutant
Bacteroides thetaiotaomicron
3.2.1.169
Y137F
site-directed mutant
Bacteroides thetaiotaomicron
3.2.1.169
Y282F
site-directed mutant
Bacteroides thetaiotaomicron
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
3.2.1.169
1,2-dideoxy-2'-methyl-alpha-D-glucopyranoso-[2,1-d]-DELTA2'-thiazoline
NAG-thiazoline
Bacteroides thetaiotaomicron
3.2.1.169
O-(2-acetamido-2-deoxy-D-glucopyranosylidene)amino-N-phenylcarbamate
PUGNAc
Bacteroides thetaiotaomicron
3.2.1.169
streptozotocin
STZ
Bacteroides thetaiotaomicron
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.2.1.169
0.016
-
4-nitrophenyl N-acetyl-beta-D-glucosaminide
D243N mutant
Bacteroides thetaiotaomicron
3.2.1.169
0.092
-
4-nitrophenyl N-acetyl-beta-D-glucosaminide
D243A mutant
Bacteroides thetaiotaomicron
3.2.1.169
0.28
-
4-nitrophenyl N-acetyl-beta-D-glucosaminide
wild-type
Bacteroides thetaiotaomicron
3.2.1.169
0.3
-
4-nitrophenyl N-acetyl-beta-D-glucosaminide
-
Bacteroides thetaiotaomicron
3.2.1.169
0.41
-
4-nitrophenyl N-acetyl-beta-D-glucosaminide
Y282F mutant
Bacteroides thetaiotaomicron
3.2.1.169
1.18
-
4-nitrophenyl N-acetyl-beta-D-glucosaminide
Y137F mutant
Bacteroides thetaiotaomicron
3.2.1.169
1.4
-
4-nitrophenyl N-acetyl-beta-D-thioglucosaminide
-
Bacteroides thetaiotaomicron
3.2.1.169
1.47
-
4-nitrophenyl N-acetyl-beta-D-glucosaminide
D242N mutant
Bacteroides thetaiotaomicron
3.2.1.169
4.4
-
4-nitrophenyl N-acetyl-beta-D-glucosaminide
D242A mutant
Bacteroides thetaiotaomicron
3.2.1.169
36
-
4-nitrophenyl N-acetyl-beta-D-glucosaminide
N372A mutant
Bacteroides thetaiotaomicron
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
3.2.1.169
additional information
Bacteroides thetaiotaomicron
enzyme cleaves 4-nitrophenyl N-acetyl-beta-D-glucosaminide from modified proteins and also cleaves 4-nitrophenyl N-acetyl-beta-D-thioglucosaminide
?
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.2.1.169
affinity chromatography using nickel-affinity chromatography followed by gel filtration
Bacteroides thetaiotaomicron
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
3.2.1.147
4-nitrophenyl 2-acetamido-2-deoxy-1-thio-beta-D-glucopyranoside + H2O
-
705869
Bacteroides thetaiotaomicron
4-nitrobenzenethiol + N-acetyl-D-glucosamine
-
-
-
?
3.2.1.169
4-methylumbelliferyl N-acetyl-beta-D-glucosaminide + H2O
-
705869
Bacteroides thetaiotaomicron
4-methylumbelliferone + N-acetyl-beta-D-glucosamine
-
-
-
?
3.2.1.169
4-methylumbelliferyl N-difluoroacetyl-beta-D-glucosaminide + H2O
-
705869
Bacteroides thetaiotaomicron
4-methylumbelliferone + N-difluoroacetyl-beta-D-glucosamine
-
-
-
?
3.2.1.169
4-methylumbelliferyl N-fluoroacetyl-beta-D-glucosaminide + H2O
-
705869
Bacteroides thetaiotaomicron
4-methylumbelliferone + N-fluoroacetyl-beta-D-glucosamine
-
-
-
?
3.2.1.169
4-methylumbelliferyl N-trifluoroacetyl-beta-D-glucosaminide + H2O
-
705869
Bacteroides thetaiotaomicron
4-methylumbelliferone + N-trifluoroacetyl-beta-D-glucosamine
-
-
-
?
3.2.1.169
4-nitrophenyl N-acetyl-beta-D-glucosaminide + H2O
-
705869
Bacteroides thetaiotaomicron
4-nitrophenol + N-acetyl-beta-D-glucosamine
-
-
-
?
3.2.1.169
4-nitrophenyl N-acetyl-beta-D-thioglucosaminide + H2O
-
705869
Bacteroides thetaiotaomicron
4-nitrophenol + N-acetyl-beta-D-thioglucosamine
-
-
-
?
3.2.1.169
additional information
enzyme cleaves 4-nitrophenyl N-acetyl-beta-D-glucosaminide from modified proteins and also cleaves 4-nitrophenyl N-acetyl-beta-D-thioglucosaminide
705869
Bacteroides thetaiotaomicron
?
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
3.2.1.169
dimer
-
Bacteroides thetaiotaomicron
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
3.2.1.169
37
-
assay at
Bacteroides thetaiotaomicron
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.2.1.147
7.4
-
assay at
Bacteroides thetaiotaomicron
3.2.1.169
6
-
bell-shaped pH-activity profile
Bacteroides thetaiotaomicron