BRENDA - Enzyme Database show
show all sequences of 4.2.2.19

Biochemical characterization of the chondroitinase B active site

Pojasek, K.; Raman, R.; Kiley, P.; Venkataraman, G.; Sasisekharan, R.; J. Biol. Chem. 277, 31179-31186 (2002)

Data extracted from this reference:

Engineering
Amino acid exchange
Commentary
Organism
E333A
PCR site-directed mutagenesis, E333 is a key residue in catalysis, reduced activity
Pedobacter heparinus
H272A
PCR site-directed mutagenesis, H272 is a key residue in catalysis, reduced activity
Pedobacter heparinus
K250A
PCR site-directed mutagenesis, inactive mutant
Pedobacter heparinus
R271A
PCR site-directed mutagenesis, R271 is a key residue in catalysis
Pedobacter heparinus
R363A
PCR site-directed mutagenesis, R363 is a key residue in catalysis, 2fold increased activity
Pedobacter heparinus
R364A
PCR site-directed mutagenesis, highly reduced activity, altered product profil, residue is involved in determination of substrate specificity
Pedobacter heparinus
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
kinetics of wild-type and mutant enzymes
Pedobacter heparinus
0.0027
-
dermatan sulfate
mutant H272A, pH 8.0, 30°C
Pedobacter heparinus
0.0028
-
dermatan sulfate
mutant E333A, pH 8.0, 30°C
Pedobacter heparinus
0.0046
-
dermatan sulfate
mutant R363A, pH 8.0, 30°C; wild-type enzyme, pH 8.0, 30°C
Pedobacter heparinus
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
additional information
no bound Ca2+
Pedobacter heparinus
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Pedobacter heparinus
Q46079
-
-
Reaction
Reaction
Commentary
Organism
Eliminative cleavage of dermatan sulfate containing (1->4)-beta-D-hexosaminyl and (1->3)-beta-D-glucurosonyl or (1->3)-alpha-L-iduronosyl linkages to disaccharides containing 4-deoxy-beta-D-gluc-4-enuronosyl groups to yield a 4,5-unsaturated dermatan-sulfate disaccharide (DeltaUA-GalNAc-4S).
acts on dermatan sulfate as sole substrate, active site structure involving Lys250, Arg271, His272, and Glu333, substrate binding mechanism involving Arg363 and Arg364
Pedobacter heparinus
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
dermatan sulfate
-
652290
Pedobacter heparinus
?
-
-
-
-
dermatan sulfate
analysis of interactions between enzyme and substrate
652290
Pedobacter heparinus
?
-
652290
Pedobacter heparinus
?
Subunits
Subunits
Commentary
Organism
More
structural analysis
Pedobacter heparinus
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Pedobacter heparinus
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
4.6
-
dermatan sulfate
mutant E333A, pH 8.0, 30°C
Pedobacter heparinus
29
-
dermatan sulfate
mutant H272A, pH 8.0, 30°C
Pedobacter heparinus
190
-
dermatan sulfate
wild-type enzyme, pH 8.0, 30°C
Pedobacter heparinus
404
-
dermatan sulfate
mutant R363A, pH 8.0, 30°C
Pedobacter heparinus
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8
-
assay at
Pedobacter heparinus
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
E333A
PCR site-directed mutagenesis, E333 is a key residue in catalysis, reduced activity
Pedobacter heparinus
H272A
PCR site-directed mutagenesis, H272 is a key residue in catalysis, reduced activity
Pedobacter heparinus
K250A
PCR site-directed mutagenesis, inactive mutant
Pedobacter heparinus
R271A
PCR site-directed mutagenesis, R271 is a key residue in catalysis
Pedobacter heparinus
R363A
PCR site-directed mutagenesis, R363 is a key residue in catalysis, 2fold increased activity
Pedobacter heparinus
R364A
PCR site-directed mutagenesis, highly reduced activity, altered product profil, residue is involved in determination of substrate specificity
Pedobacter heparinus
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
kinetics of wild-type and mutant enzymes
Pedobacter heparinus
0.0027
-
dermatan sulfate
mutant H272A, pH 8.0, 30°C
Pedobacter heparinus
0.0028
-
dermatan sulfate
mutant E333A, pH 8.0, 30°C
Pedobacter heparinus
0.0046
-
dermatan sulfate
mutant R363A, pH 8.0, 30°C; wild-type enzyme, pH 8.0, 30°C
Pedobacter heparinus
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
additional information
no bound Ca2+
Pedobacter heparinus
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
dermatan sulfate
-
652290
Pedobacter heparinus
?
-
-
-
-
dermatan sulfate
analysis of interactions between enzyme and substrate
652290
Pedobacter heparinus
?
-
652290
Pedobacter heparinus
?
Subunits (protein specific)
Subunits
Commentary
Organism
More
structural analysis
Pedobacter heparinus
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Pedobacter heparinus
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
4.6
-
dermatan sulfate
mutant E333A, pH 8.0, 30°C
Pedobacter heparinus
29
-
dermatan sulfate
mutant H272A, pH 8.0, 30°C
Pedobacter heparinus
190
-
dermatan sulfate
wild-type enzyme, pH 8.0, 30°C
Pedobacter heparinus
404
-
dermatan sulfate
mutant R363A, pH 8.0, 30°C
Pedobacter heparinus
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8
-
assay at
Pedobacter heparinus
Other publictions for EC 4.2.2.19
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)
747452
Wu
Establishment of chondroitin ...
Pedobacter heparinus, Pedobacter heparinus DSM 2366
Carbohydr. Polym.
144
338-345
2016
-
1
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2
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692532
Volpi
Structural characterization an ...
Pedobacter heparinus
Glycobiology
19
356-367
2008
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6
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1
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1
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1
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1
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677536
Oguma
Analytical method for the dete ...
Pedobacter heparinus
Anal. Biochem.
368
79-86
2007
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3
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1
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1
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677405
Makovitzky
Topooptical investigations and ...
Pedobacter heparinus
Acta Histochem.
108
193-196
2006
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-
-
-
-
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1
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1
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1
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680938
Nakamura
Glycosaminoglycan degradation ...
Pedobacter heparinus
J. Biomed. Mater. Res. A
77
478-486
2006
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3
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1
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1
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652553
Michel
The structure of chondroitin B ...
Pedobacter heparinus
J. Biol. Chem.
279
32882-32896
2004
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1
6
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1
2
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1
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2
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1
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1
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1
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1
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2
1
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1
6
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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
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1
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652290
Pojasek
Biochemical characterization o ...
Pedobacter heparinus
J. Biol. Chem.
277
31179-31186
2002
-
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6
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4
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1
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2
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1
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2
1
1
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4
1
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6
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4
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1
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2
1
1
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4
1
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652640
Suzuki
Inhibition of hyaluronidases a ...
Pedobacter heparinus
J. Enzyme Inhib. Med. Chem.
17
183-186
2002
-
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25
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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26
2
1
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1
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1
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649575
Pojasek
Recombinant expression, purifi ...
Pedobacter heparinus
Biochem. Biophys. Res. Commun.
286
343-351
2001
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1
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2
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1
1
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651192
Denholm
Anti-tumor activities of chond ...
Homo sapiens
Eur. J. Pharmacol.
416
213-221
2001
-
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2
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649356
Tkalec
Isolation and expression in Es ...
Pedobacter heparinus
Appl. Environ. Microbiol.
66
29-35
2000
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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3
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1
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649099
Li
Crystallization and preliminar ...
Pedobacter heparinus
Acta Crystallogr. Sect. D
55
1055-1057
1999
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1
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652806
Huang
Crystal structure of chondroit ...
Pedobacter heparinus
J. Mol. Biol.
294
1257-1269
1999
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1
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1
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3
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1
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1
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34100
Gu
Purification, characterization ...
Pedobacter heparinus
Biochem. J.
312
569-577
1995
-
-
-
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1
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1
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2
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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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650773
Ototani
Purification of chondroitinase ...
Pedobacter heparinus
Carbohydr. Res.
70
295-306
1979
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