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Literature summary for 4.2.2.21 extracted from

  • Prabhakar, V.; Capila, I.; Soundararajan, V.; Raman, R.; Sasisekharan, R.
    Recombinant expression, purification, and biochemical characterization of chondroitinase ABC II from Proteus vulgaris (2009), J. Biol. Chem., 284, 974-982.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Proteus vulgaris

Protein Variants

Protein Variants Comment Organism
E608A the mutant does not show detectable activity on either chondroitin 6-sulfate or dermatan sulfate Proteus vulgaris
H452A the mutant shows similar Km values to the wild type enzyme but a 100fold reduced catalytic efficiency Proteus vulgaris
H453A the mutant does not show detectable activity on either chondroitin 6-sulfate or dermatan sulfate Proteus vulgaris
H456A the mutant shows about 3fold reduction in catalytic efficiency Proteus vulgaris
R513A the mutant does not show detectable activity on either chondroitin 6-sulfate or dermatan sulfate Proteus vulgaris
Y460A the mutant does not show detectable activity on either chondroitin 6-sulfate or dermatan sulfate Proteus vulgaris

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0076
-
dermatan sulfate mutant enzyme H456A, in 50mM Tris/HCl, pH 8.0, at 37°C Proteus vulgaris
0.0077
-
chondroitin 6-sulfate mutant enzyme H452A, in 50mM Tris/HCl, pH 8.0, at 37°C Proteus vulgaris
0.0091
-
dermatan sulfate mutant enzyme H452A, in 50mM Tris/HCl, pH 8.0, at 37°C Proteus vulgaris
0.0098
-
chondroitin 6-sulfate wild type enzyme, in 50mM Tris/HCl, pH 8.0, at 37°C Proteus vulgaris
0.0104
-
chondroitin 6-sulfate mutant enzyme H456A, in 50mM Tris/HCl, pH 8.0, at 37°C Proteus vulgaris
0.0161
-
chondroitin 4-sulfate wild type enzyme, in 50mM Tris/HCl, pH 8.0, at 37°C Proteus vulgaris
0.0192
-
dermatan sulfate wild type enzyme, in 50mM Tris/HCl, pH 8.0, at 37°C Proteus vulgaris

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
100000
-
SDS-PAGE Proteus vulgaris

Organism

Organism UniProt Comment Textmining
Proteus vulgaris
-
strain ATCC 6896
-

Purification (Commentary)

Purification (Comment) Organism
Hi-Trap Ni-chelate column chromatography, gel filtration Proteus vulgaris

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
chondroitin
-
Proteus vulgaris ?
-
?
chondroitin 4-sulfate
-
Proteus vulgaris unsaturated chondroitin 4-sulfate disaccharide the major product ?
chondroitin 6-sulfate
-
Proteus vulgaris ?
-
?
chondroitin sulfate D
-
Proteus vulgaris ?
-
?
chondroitin sulfate E
-
Proteus vulgaris ?
-
?
dermatan sulfate
-
Proteus vulgaris 2-acetamido-2-deoxy-3-O-(3-D-gluco-4-enepyranosyluronic acid)-4-O-sulfo-D-galactose + 2-acetamido-2-deoxy-3-O-(3-D-gluco-4-enepyranosyluronic acid)-6-O-sulfo-D-galactose + 2-acetamido-2-deoxy-3-O-(3-D-gluco-4-enepyranosyluronic acid)-4,6-O-bis-sulfo-D-galactose
-
?
additional information cABCII is an exolytic enzyme that cleaves the substrate only at the nonreducing end of the polysaccharide Proteus vulgaris ?
-
?
additional information the recombinant protein with the sequence discrepancies is unable to effectively process galactosaminoglycan substrates, the enzyme does not react with hyaluronan, heparin/heparan sulfate, and keratan sulfate Proteus vulgaris ?
-
?

Synonyms

Synonyms Comment Organism
cABCII isoform Proteus vulgaris
chondroitinase ABC II
-
Proteus vulgaris

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.072
-
dermatan sulfate mutant enzyme H452A, in 50mM Tris/HCl, pH 8.0, at 37°C Proteus vulgaris
0.17
-
chondroitin 6-sulfate mutant enzyme H452A, in 50mM Tris/HCl, pH 8.0, at 37°C Proteus vulgaris
2.4
-
dermatan sulfate mutant enzyme H456A, in 50mM Tris/HCl, pH 8.0, at 37°C Proteus vulgaris
7.13
-
chondroitin 6-sulfate mutant enzyme H456A, in 50mM Tris/HCl, pH 8.0, at 37°C Proteus vulgaris
16.67
-
chondroitin 4-sulfate wild type enzyme, in 50mM Tris/HCl, pH 8.0, at 37°C Proteus vulgaris
19.17
-
dermatan sulfate wild type enzyme, in 50mM Tris/HCl, pH 8.0, at 37°C Proteus vulgaris
21.67
-
chondroitin 6-sulfate wild type enzyme, in 50mM Tris/HCl, pH 8.0, at 37°C Proteus vulgaris

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
8.3
-
dermatan sulfate mutant enzyme H452A, in 50mM Tris/HCl, pH 8.0, at 37°C Proteus vulgaris
23.3
-
chondroitin 6-sulfate mutant enzyme H452A, in 50mM Tris/HCl, pH 8.0, at 37°C Proteus vulgaris
316.7
-
dermatan sulfate mutant enzyme H456A, in 50mM Tris/HCl, pH 8.0, at 37°C Proteus vulgaris
683.3
-
chondroitin 6-sulfate mutant enzyme H456A, in 50mM Tris/HCl, pH 8.0, at 37°C Proteus vulgaris
1000
-
dermatan sulfate wild type enzyme, in 50mM Tris/HCl, pH 8.0, at 37°C Proteus vulgaris
1000
-
chondroitin 4-sulfate wild type enzyme, in 50mM Tris/HCl, pH 8.0, at 37°C Proteus vulgaris
2200
-
chondroitin 6-sulfate wild type enzyme, in 50mM Tris/HCl, pH 8.0, at 37°C Proteus vulgaris