Information on EC 3.2.1.102 - blood-group-substance endo-1,4-beta-galactosidase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.2.1.102
-
RECOMMENDED NAME
GeneOntology No.
blood-group-substance endo-1,4-beta-galactosidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Endohydrolysis of (1->4)-beta-D-galactosidic linkages in blood group A and B substances
show the reaction diagram
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-
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-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of O-glycosyl bond
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-
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SYSTEMATIC NAME
IUBMB Comments
blood-group-substance 4-beta-D-galactanohydrolase
Hydrolyses the 1,4-beta-D-galactosyl linkages adjacent to a 1,3-alpha-D-galactosyl or N-acetylgalactosaminyl residues and a 1,2-alpha-D-fucosyl residue.
CAS REGISTRY NUMBER
COMMENTARY hide
52720-51-1
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain ATCC 10543, strain 13, gene eabC
SwissProt
Manually annotated by BRENDA team
strain SP3-BS71
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-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
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endo-beta-galactosidase C overexpression results in accelerated proliferation of mouse NIH3T3 fibroblasts
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2,4-dinitrophenyl 2-acetamido-2-deoxy-D-galactopyranosyl-(1,3)-[alpha-L-fucopyranosyl-(1,2)]-beta-D-galactopyranoside + H2O
?
show the reaction diagram
-
-
-
-
?
alpha-galactosyl antigen + H2O
?
show the reaction diagram
-
erythrocyte from pig
-
-
?
blood group A glycoconjugate + H2O
?
show the reaction diagram
blood group B glycoconjugate + H2O
?
show the reaction diagram
blood group O glycopeptide + H2O
GlcNAcbeta1-3Gal-ol + Galbeta1-4GlcNAcbeta1-3Gal-ol + Fucalpha1-2Galbeta1-4GlcNAcbeta1-3Gal-ol + GlcNAcbeta1-6[GlcNAcbeta1-3]Galbeta1-4GlcNAcbeta1-3Gal-ol + Galbeta1-4GlcNAcbeta1-6[GlcNAcbeta1-3]Galbeta1-4GlcNAcbeta1-3Gal-ol + GlcNAcbeta1-6[Galbeta1-4GlcNAcbeta1-3]Galbeta1-4GlcNAcbeta1-3Gal-ol + Galbeta1-4GlcNAcbeta1-6[Galbeta1-4GlcNAcbeta1-3]Galbeta1-4GlcNAcbeta1-3Gal-ol + Fucalpha1-2Galbeta1-4GlcNAcbeta1-6[GlcNAcbeta1-3]Galbeta1-4GlcNAcbeta1-3Gal-ol + Fucalpha1-2Galbeta1-4GlcNAcbeta1-6[Galbeta1-4GlcNAcbeta1-3]Galbeta1-4GlcNAcbeta1-3Gal-ol + Fucalpha1-2Galbeta1-4GlcNAcbeta1-6[Fucalpha1-2Galbeta1-4GlcNAcbeta1-3]Galbeta1-4GlcNAcbeta1-3Gal-ol
show the reaction diagram
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desialylated human blood group O erythrocyte glycopeptide
characterization of the oligosaccharides
?
blood group substance A + H2O
GalNAcalpha1-3(Fucalpha1-2)Gal + Galalpha1-3(Fucalpha1-2)Gal + ?
show the reaction diagram
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degradation of blood type A determinant
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?
blood group substance B + H2O
?
show the reaction diagram
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degradation of blood type B determinant
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-
?
Galalpha(1-3)[Fucalpha(1-2)]Galbeta(1-4)GlcNAcbeta-CH2-CH2-N3 + H2O
?
show the reaction diagram
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analog of blood group B antigen
-
-
?
Galalpha(1-3)[Fucalpha(1-2)]Galbeta(1-4)GlcNAcbeta-R + H2O
Galalpha(1-3)[Fucalpha(1-2)]Gal + GlcNAcbeta-R
show the reaction diagram
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analog of blood group B antigen, digested more effciently than the blood group A analog
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-
?
Galalpha1-3(Fucalpha1-2)Galbeta1-4(Fucalpha1-3)GlcNAcbeta1-3-hexene-1,2,5,6-tetrol + H2O
?
show the reaction diagram
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-
-
-
?
Galalpha1-3(Fucalpha1-2)Galbeta1-4GlcNAcbeta1-3-hexene-1,2,5,6-tetrol + H2O
?
show the reaction diagram
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-
-
-
?
Galalpha1-3Galbeta1-4GlcNAcbeta1-3Galbeta1-4Glc + H2O
Galalpha1-3Galbeta + GlcNAcbeta1-3Galbeta1-4Glc
show the reaction diagram
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-
-
-
?
Galbeta1-3GlcNAcbeta1-3Galbeta1-4Glcbeta-p-nitrophenol + H2O
Galbeta1-3GlcNAcbeta1-3Galbeta + Glcbeta-p-nitrophenol
show the reaction diagram
Galbeta1-4GlcNAcbeta1-3Galbeta1-4Glcbeta-p-nitrophenol + H2O
Galbeta1-4GlcNAcbeta1-3Galbeta + Glcbeta-p-nitrophenol
show the reaction diagram
Galbeta1-4GlcNAcbeta1-3Galbeta1-4GlcNAcbeta-p-nitrophenol + H2O
Galbeta1-4GlcNAcbeta1-3Galbeta + GlcNAcbeta-p-nitrophenol
show the reaction diagram
Galbeta1-6GlcNAcbeta1-3Galbeta1-4Glcbeta-p-nitrophenol + H2O
Galbeta1-6GlcNAcbeta1-3Galbeta + Glcbeta-p-nitrophenol
show the reaction diagram
GalNAcalpha(1-3)[Fucalpha(1-2)]Galbeta(1-4)GlcNAcbeta-CH2-CH2-N3 + H2O
?
show the reaction diagram
-
analog of blood group A antigen
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-
?
GalNAcalpha(1-3)[Fucalpha(1-2)]Galbeta(1-4)GlcNAcbeta-R + H2O
GalNAcalpha(1-3)[Fucalpha(1-2)]Gal + GlcNAcbeta-R
show the reaction diagram
-
analog of blood group A antigen
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?
GalNAcalpha1-3(Fucalpha1-2)Galbeta1-4Glc + H2O
?
show the reaction diagram
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?
GalNAcalpha1-3(Fucalpha1-2)Galbeta1-4GlcNAcbeta1-3-hexene-1,2,5,6-tetrol + H2O
?
show the reaction diagram
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-
-
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?
GlcNAcbeta1-3Galbeta1-4Glc-p-nitrophenol + H2O
GlcNAcbeta1-3Galbeta + Glc-p-nitrophenol
show the reaction diagram
GlcNAcbeta1-3Galbeta1-4GlcNAcbeta-p-nitrophenol + H2O
GlcNAcbeta1-3Galbeta + GlcNAcbeta-p-nitrophenol
show the reaction diagram
GlcNAcbeta1-3Galbeta1-4GlcNAcbeta-p-nitrophenol + H2O
GlcNAcbeta1-3Galbeta1-4GlcNAcbeta1-3Galbeta1-4GlcNAcbeta-p-nitrophenol + GlcNAcbeta1-3Galbeta1-4GlcNAcbeta1-3Galbeta1-4GlcNAcbeta1-3Galbeta1-4GlcNAcbeta-p-nitrophenol
show the reaction diagram
keratan sulfate + H2O
?
show the reaction diagram
lactosaminoglycan + H2O
small oligosaccharides + residual glycopeptide
show the reaction diagram
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-
-
?
LewisY tetrasaccharide + H2O
?
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
blood group A glycoconjugate + H2O
?
show the reaction diagram
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-
-
-
?
blood group B glycoconjugate + H2O
?
show the reaction diagram
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-
-
-
?
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Ag2SO4
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3 mM, 82% inhibition
Cys
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3.3 mM, 15 min at 37C, 52% inhibition
HgCl2
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3 mM, 98.6% inhibition
PCMB
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3.3 mM, 15 min at 37C, 88% inhibition
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.064 - 0.61
2,4-dinitrophenyl 2-acetamido-2-deoxy-D-galactopyranosyl-(1,3)-[alpha-L-fucopyranosyl-(1,2)]-beta-D-galactopyranoside
0.44
blood group A glycoconjugate
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at 37C
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0.57
blood group B glycoconjugate
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at 37C
0.67
Galalpha1-3(Fucalpha1-2)Galbeta1-4(Fucalpha1-3)GlcNAcbeta1-3-hexene-1,2,5,6-tetrol
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0.36
Galalpha1-3(Fucalpha1-2)Galbeta1-4GlcNAcbeta1-3-hexene-1,2,5,6-tetrol
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-
0.13
GalNAcalpha1-3(Fucalpha1-2)Galbeta1-4Glc
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0.068
GalNAcalpha1-3(Fucalpha1-2)Galbeta1-4GlcNAcbeta1-3-hexene-1,2,5,6-tetrol
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.17 - 1.75
2,4-dinitrophenyl 2-acetamido-2-deoxy-D-galactopyranosyl-(1,3)-[alpha-L-fucopyranosyl-(1,2)]-beta-D-galactopyranoside
4.67
blood group A glycoconjugate
Streptococcus pneumoniae
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at 37C
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11.17
blood group B glycoconjugate
Streptococcus pneumoniae
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at 37C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5.33 - 26.67
2,4-dinitrophenyl 2-acetamido-2-deoxy-D-galactopyranosyl-(1,3)-[alpha-L-fucopyranosyl-(1,2)]-beta-D-galactopyranoside
8847
10.67
blood group A glycoconjugate
Streptococcus pneumoniae
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at 37C
35655
20
blood group B glycoconjugate
Streptococcus pneumoniae
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at 37C
35666
0.43
LewisY tetrasaccharide
Streptococcus pneumoniae
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at 37C
156524
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1500
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still acitve after 500-fold dilution of the enzyme
2894
purified native enzyme
additional information
-
2-3fold preference for blood group A antigen
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 7.5
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pH 5.0: about 20% of maximal activity, pH 7.5: about 25% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
88000
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SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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the enzyme contains a central catalytic TIM barrel domain, the GH98 catalytic domain, and two other domains, domain organization, overview
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop vapor diffusion method, 18C
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hanging drop vapor diffusion method, using 17% (v/v) polyethylene glycol 3350, 0.2 M ammonium sulfate, and 0.1 M sodium acetate trihydrate, pH 4.8
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
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stable at 4C
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
HisTrap column chromatography and MonoQ column chromatography
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immobilized metal affinity chromatography
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native enzyme from strain ATCC 10543, by gel filtration, ion exchange, adsorption, and concanavalin A affinity chromatography, and another step of ion exchange chromatography, 2630fold to homogeneity, recombinant His-tagged enzyme by nickel affinity chromatography
Ni-NTA column chromatography and Sephacryl S-200 gel filtration
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nickel-nitrilotriacetic acid column
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis
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expressed in Escherichia coli BL21(DE3) cells
expressed in NIH3T3 cells
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expression in Escherichia coli
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expression in Escherichia coli BL-21
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expression in Mus musculus
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gene eabC, DNA and amino acid sequence determination and analysis, functional overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D429A
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the mutants shows reduced activity compared to the wild type enzyme
D453A
-
the mutation results in complete loss of activity
E354A
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the activity of the E354A mutant with nonactivated natural substrates is 1100fold lower than that of the wild type enzyme, while its activity is only 10fold lower when assayed with 2,4-dinitrophenyl-beta-A-trisaccharide
E467A
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the mutants shows slightly reduced activity compared to the wild type enzyme
E506A
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the mutation results in complete loss of activity
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
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endo-beta-galactosidase-Griffonia-simplicifolia-agglutinin-II-staining procedure into carbohydrate histochemistry of human tissues. The procedure may provide a powerful tool for analysing the backbone structures carrying the blood-group-related antigens in tissue sections
medicine