Information on EC 2.4.1.79 - globotriaosylceramide 3-beta-N-acetylgalactosaminyltransferase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY
2.4.1.79
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RECOMMENDED NAME
GeneOntology No.
globotriaosylceramide 3-beta-N-acetylgalactosaminyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
UDP-N-acetyl-alpha-D-galactosamine + alpha-D-galactosyl-(1->4)-beta-D-galactosyl-(1->4)-beta-D-glucosyl-(1<->1)-ceramide = UDP + N-acetyl-beta-D-galactosaminyl-(1->3)-alpha-D-galactosyl-(1->4)-beta-D-galactosyl-(1->4)-beta-D-glucosyl-(1<->1)-ceramide
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hexosyl group transfer
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PATHWAY
KEGG Link
MetaCyc Link
Glycosphingolipid biosynthesis - globo series
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Metabolic pathways
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SYSTEMATIC NAME
IUBMB Comments
UDP-N-acetyl-D-galactosamine:alpha-D-galactosyl-(1->4)-beta-D-galactosyl-(1->4)-beta-D-glucosyl-(1<->1)-ceramide III3-beta-N-acetyl-D-galactosaminyltransferase
Globoside is a neutral glycosphingolipid in human erythrocytes and has blood-group-P-antigen activity [4]. The enzyme requires a divalent cation for activity, with Mn2+ required for maximal activity [3]. UDP-GalNAc is the only sugar donor that is used efficiently by the enzyme: UDP-Gal and UDP-GlcNAc result in very low enzyme activity [3]. Lactosylceramide, globoside and gangliosides GM3 and GD3 are not substrates [4]. For explanation of the superscripted '3' in the systematic name, see {iupac/misc/glylp#5.3::GL-5.3.4}.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
acetylgalactosaminyltransferase, uridine diphosphoacetylgalactosamine-galactosylgalactosylglucosylceramide
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acetylgalactosaminyltransferase, uridine diphosphoacetylgalactosamine-glycosphingolipid
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beta(1->3) N-acetylgalactosaminyltransferase
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beta1,3GalNAc-T
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galactosylgalactosylglucosylceramide beta-D-acetylgalactosaminyltransferase
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GalNAc transferase
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globoside N-acetylgalactosaminyltransferase
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globoside synthetase
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glycosphingolipid beta-N-acetylgalactosaminyltransferase
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UDP-N-acetylgalactosamine:globotriaosylceramide 3-beta-N-acetylgalactosaminyltransferase
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UDP-N-acetylgalactosamine:globotriaosylceramide beta-3-N-acetylgalactosaminyltransferase
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CAS REGISTRY NUMBER
COMMENTARY
62213-46-1
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83215-90-1
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ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
embryo
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Manually annotated by BRENDA team
Haemophilus influenzae RD
strain Rd
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Manually annotated by BRENDA team
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Co2+
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46% of the activation with Mn2+
Co2+
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43% of the activation with Mn2+
Fe2+
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19% of the activation with Mn2+
Li+
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11% of the activation with Mn2+
Mg2+
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18% as effective as Mn2+ in activation
Mn2+
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required for maximal activity, 4 mM
Mn2+
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optimal concentration: 10 mM; required
Mn2+
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required
Mn2+
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required
additional information
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Mg2+ cannot replace Mn2+
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
4-hydroxymercuribenzoate
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0.5 mM, complete inhibition
EDTA
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25 mM, 99% inhibition
Globotetraosylceramide
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GSH
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2 mM, 17% inhibition
II3-alpha-N-Acetylneuraminyl-lactosylceramide
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iodoacetamide
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1 mM, 65% inhibition
NaN3
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2 mM, 38% inhibition
NEM
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1 mM, 67% inhibition
Sodium deoxycholate
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UDP
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competitive to UDP-N-acetylgalactosamine, non-competitive to globotriaosylceramide
iodoacetic acid
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1 mM, 91% inhibition
additional information
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no effect: lactosylceramide
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ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
dithiothreitol
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10 mM, stimulation to 120% of the original activity
Myrj 59
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activation is half as effective as with taurocholate
Nonionic detergents
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activation, e.g. Triton X-100 or CF-54
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Nonionic detergents
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activation, e.g. Triton X-100 or CF-54
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sodium taurocholate
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required for optimal activity
sodium taurodeoxycholate
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greatest activation, 1 mg/ml
Triton CF-54
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activates
Triton X100
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activates
Tween 20
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activation is half as effective as with taurocholate
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0025
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D-galactosyl-1,4-D-galactosyl-1,4-D-glucosylceramide
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0.14
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D-galactosyl-1,4-D-galactosyl-1,4-D-glucosylceramide
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1.7
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D-galactosyl-1,4-D-galactosyl-1,4-D-glucosylceramide
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0.42
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globotriosylceramide
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0.14
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UDP-N-acetyl-D-galactosamine
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0.2
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UDP-N-acetylgalactosamine
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0.23
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UDP-N-acetylgalactosamine
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0.014
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UDP-N-acetylglucosamine
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SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.5
8
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maximal activity in 2-(N-morpholino)ethanesulfonic acid
6.5
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7.2
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pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.5
8
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about 50% of maximal activity at pH 5.5 and 8.0
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
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assay at
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
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MW of the bifunctional beta(1,3)-N-acetylgalactosaminyltransferase/UDP-N-acetylglucosamine C4 epimerase fusion protein is 70000 Da
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
diluted enzyme solutions, below 0.1 mg protein/ml, unstable
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Stable to repeated freeze-thawing
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STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-80°C, above 0.1 mg protein/ml in 25 mM sodium cacodylate buffer, pH 6.9, 1% Triton X-100 and 20% glycerol, stable for at least 3 months
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Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
one-step purification of the bifunctional beta(1->3) N-acetylgalactosaminyltransferase/UDP-N-acetylglucosamine C4 epimerase fusion protein
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ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
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preparation of a bifunctional beta(1->3) N-acetylgalactosaminyltransferase/UDP-N-acetylglucosamine C4 epimerase fusion protein by in-frame linking of the Plesiomonas shigelloides wbgU gene downstream to the Haemophilus influenzae lgtD gene through a five-residue peptide linker (Thr-Gly-Gly-Gly-Gly). The enzyme is expressed in Escherichia coli BL21 at a relatively high level
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
synthesis
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synthesis of globoside and isogloboside tetrasaccharides by using beta(1->3) N-acetylgalactosaminyltransferase/UDP-N-acetylglucosamine C4 epimerase fusion protein
synthesis
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synthesis of globotetraose (GalNAcb-3Gala-4Galb-4Glc)by the high cell density culture of an Escherichia coli strain over-expressing the Neisseria meningitidis lgtC gene for alpha-1,4-Gal transferase. The strain which is devoid of both alpha and beta galactosidase activity is fed with glycerol as the energy and carbon source and with lactose as precursor for globotriose synthesis. After complete exhaustion of lactose, globotriose could serve as an alternative acceptor for LgtC and the formation of a series of polygalactosylated compounds is observed. The system is extended to the synthesis of globotetraose (GalNAcbeta-3Gala-4Galbeta-4Glc) by overexpressing two additional genes: lgtD from Haemophilus influenzae Rd which encodes a beta-1,3-GalNAc transferase and wbpP from Pseudomonas aeruginosa which encodes a UDP-GalNAc C4 epimerase. Globotetraose could also be produced from exogenous globotriose which is actively taken up by the cells
synthesis
Haemophilus influenzae RD
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synthesis of globotetraose (GalNAcb-3Gala-4Galb-4Glc)by the high cell density culture of an Escherichia coli strain over-expressing the Neisseria meningitidis lgtC gene for alpha-1,4-Gal transferase. The strain which is devoid of both alpha and beta galactosidase activity is fed with glycerol as the energy and carbon source and with lactose as precursor for globotriose synthesis. After complete exhaustion of lactose, globotriose could serve as an alternative acceptor for LgtC and the formation of a series of polygalactosylated compounds is observed. The system is extended to the synthesis of globotetraose (GalNAcbeta-3Gala-4Galbeta-4Glc) by overexpressing two additional genes: lgtD from Haemophilus influenzae Rd which encodes a beta-1,3-GalNAc transferase and wbpP from Pseudomonas aeruginosa which encodes a UDP-GalNAc C4 epimerase. Globotetraose could also be produced from exogenous globotriose which is actively taken up by the cells
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