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2.4.1.87: N-acetyllactosaminide 3-alpha-galactosyltransferase

This is an abbreviated version!
For detailed information about N-acetyllactosaminide 3-alpha-galactosyltransferase, go to the full flat file.

Word Map on EC 2.4.1.87

Reaction

UDP-alpha-D-galactose
+
beta-D-galactosyl-(1->4)-beta-N-acetyl-D-glucosaminyl-R
=
UDP
+
alpha-D-galactosyl-(1->3)-beta-D-galactosyl-(1->4)-beta-N-acetylglucosaminyl-R

Synonyms

alpha(1,3)GT, alpha-1,3-galactosyltransferase, alpha-1,3galactosyltransferase, alpha-D-galactosyltransferase, alpha-galactosyltransferase, alpha1,3-galactosyltransferase, alpha1,3-GalT, alpha1,3galactosyltransferase, alpha1,3GalT, alpha1,3GT, alpha3-galactosyltransferase, alpha3Gal-T, alpha3GalT, alpha3GT, beta-D-galactosyl-N-acetylglucosaminylglycopeptide alpha-1,3-galactosyltransferase, blood group glycosyltransferase, EC 2.4.1.124, EC 2.4.1.151, galactosyltransferase, uridine diphosphogalactose-acetyllactosamine, galactosyltransferase, uridine diphosphogalactose-acetyllactosamine alpha1-->3-, galactosyltransferase, uridine diphosphogalactose-galactosylacetylglucosamine alpha-1,3-, galactosyltransferase, uridine diphosphogalactose-galactosylacetylglucosaminylgalactosylglucosylceramide, GALT, GGTA1, glucosaminylglycopeptide alpha-1,3-galactosyltransferase, GTB, iGb3 synthase, iGb3S, N-acetyllactoseaminide 3-alpha-D-galactosyltransferase, one, three-galactosyltransferase, Otg1p, Otg2p, Otg3p, UDP-D-Gal(1,4)-D-GlcNAc alpha(1,3)-galactosyltransferase, UDP-Gal:N-acetyllactoseaminide alpha(1,3)-galactosyltransferase, UDP-galactose beta galactosyl alpha1,3-galactosyltransferase, UDP-galactose beta-galactosyl alpha-1,3-galactosyltransferase, UDP-galactose-acetyllactosamine alpha-D-galactosyltransferase, UDP-galactose-acetyllactoseamine alpha-D-galactosyltransferase, UDP-galactose:beta-D-galactosyl-beta-1,4-N-acetyl-D-glucosaminyl-glycopeptide alpha-1,3-D-galactosyltransferase, UDP-galactose:N-acetyllactoseaminide 3-alpha-D-galactosyltransferase, UDP-GalN-acetyllactoseaminide alpha-1,3-D-UDP-GalGalbeta1-->4GlcNAC-R alpha1-->3-galactosyltransferase, UDPgalactose:beta-D-galactosyl-beta-1,4-N-acetyl-D-glucosaminyl-glycopeptide alpha-1,3-D-galactosyltransferase, uridine diphosphogalactose-acetyllactosamine alpha1-->3-galactosyltransferase, uridine diphosphogalactose-galactosylacetylglucosaminylgalactosylglucosylceramide galactosyltransferase, WciN, wclR

ECTree

     2 Transferases
         2.4 Glycosyltransferases
             2.4.1 Hexosyltransferases
                2.4.1.87 N-acetyllactosaminide 3-alpha-galactosyltransferase

Engineering

Engineering on EC 2.4.1.87 - N-acetyllactosaminide 3-alpha-galactosyltransferase

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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D316E
D316N
E317A
relative activity of mutant protein: 0.1% (compared to wild-type 100%)
E317A/E317Q
-
site-directed mutagenesis, substrate binding compared to wild-type
E317C
relative activity of mutant protein: 0.8% (compared to wild-type 100%)
E317D
relative activity of mutant protein: 0.4% (compared to wild-type 100%)
E317H
relative activity of mutant protein: 0.1% (compared to wild-type 100%)
E317Q
H280A/A281A/A282A
mutant AAA, 20 or 200 mM lactose as acceptor, 14 and 41% galactosyltransferase activity (UDP-galactose as donor), 4 and 6% N-acetylgalactosamine transferase activity (UDP-N-acetylgalactosamine as donor), 37°C, MnCl2, 25 mM Tris-HCl, pH 7.0
H280A/A281G/A282G
mutant AGG, 20 or 200 mM lactose as acceptor, 0.4 and 1% galactosyltransferase activity (UDP-galactose as donor), 0.3 and 2% N-acetylgalactosamine transferase activity (UDP-N-acetylgalactosamine as donor), 37°C, MnCl2, 25 mM Tris-HCl, pH 7.0, among the mutants with highest N-acetylgalactosamnine transferase activity and ability to transfer C2-modified sugars
H280A/A281G/A282G/L283I
AGGI-mutant, instable, leucin 283 is important for enzyme stability
H280A/A281G/A282G/L283L
AGGL-mutant, higher N-acetylgalactosamine transferase activity than galactosyltransferase activity, leucin 283 is important for enzyme stability
H280A/A281G/A282G/L283V
AGGV-mutant, instable, leucin 283 is important for enzyme stability
H280G/A281A/A282A
protein unstable, precipitating during purification
H280G/A281A/A282G
mutant GAG, 20 or 200 mM lactose as acceptor, 5 and 27% galactosyltransferase activity (UDP-galactose as donor), 4 and 6% N-acetylgalactosamine transferase activity (UDP-N-acetylgalactosamine as donor), 37°C, MnCl2, 25 mM Tris-HCl, pH 7.0
H280G/A281G/A282G/L283I
GGGI-mutant, instable, leucin 283 is important for enzyme stability
H280G/A281G/A282G/L283L
GGGL-mutant, higher N-acetylgalactosamine transferase activity than galactosyltransferase activity, leucin 283 is important for enzyme stability
H280G/A281G/A282G/L283V
GGGV-mutant, instable, leucin 283 is important for enzyme stability
H280L/A281G/A282G
mutant LGG, 20 or 200 mM lactose as acceptor, 2 and 1% galactosyltransferase activity (UDP-galactose as donor), 3 and 2% N-acetylgalactosamine transferase activity (UDP-N-acetylgalactosamine as donor), 37°C, MnCl2, 25 mM Tris-HCl, pH 7.0
H280Q
80fold reduction in catalytic activity
H280R/A281A/A282A/L283I
RAAI-mutant, no catalytic activity, crystallisation
H280S/A281G/A282G
mutant SGG, 20 or 200 mM lactose as acceptor, 5 and 5% galactosyltransferase activity (UDP-galactose as donor), 2 and 11% N-acetylgalactosamine transferase activity (UDP-N-acetylgalactosamine as donor), 37°C, MnCl2, 25 mM Tris-HCl, pH 7.0, among the mutants with highest GalNac transferase activity and ability to transfer C2-modified sugars
H280T/A281G/A282G
mutant TGG, 20 or 200 mM lactose as acceptor, 3 and 15% galactosyltransferase activity (UDP-galactose as donor), 1 and 6% N-acetylgalactosamine transferase activity (UDP-N-acetylgalactosamine as donor), 37°C, MnCl2, 25 mM Tris-HCl, pH 7.0
H280V/A281G/A282G
mutant VGG, 20 or 200 mM lactose as acceptor, 4 and 16% galactosyltransferase activity (UDP-galactose as donor), 0.5 and 3% N-acetylgalactosamine transferase activity (UDP-N-acetylgalactosamine as donor), 37°C, MnCl2, 25 mM Tris-HCl, pH 7.0
H315Q
mutant protein shows modest changes in kinetic parameters for lactose
H315R
mutant shows a major losss in catalytic activity arising from a 500fold reduction in kcat
H319A
mutant shows a 2fold reduction for kcat
H319E
mutant is inactive
H319Y
mutant is inactive
Q247E
50fold reduction in turnover number for transferase reaction, unchanged hydrolase activity, Km-values for UDP-galactose and lactose are similar to the wild-type values
R365K
mutation reduces turnover-number for UDP-galactose hydrolysis about 10fold but has no significant effect on the affinity for UDP-galactose
S318A
mutant shows a 10fold reduction for kcat
W249G
-
1.95fold increase in Km-value for UDP-galactose in galactosyltransferase reaction, turnover number of hydrolase reaction is identical to wild-type value, 2.1fold increase in Km-value for UDP-galactose in hydrolase reaction. Mutation does not affect the overall structure of the enzyme or its interactions with ligands
W250F
-
2.6fold increase in turnover number of galactosyltransferase reaction, 9.3fold increase in Km-value for UDP-galactose in galactosyltransferase reaction, 1.1fold decrease in Km-value for lactose, 1.4fold increase in turnover number of hydrolase reaction, 1.5fold increase in Km-value for UDP-galactose in hydrolase reaction
W250Y
-
1.2fold increase in turnover number of galactosyltransferase reaction, 5.2fold increase in Km-value for UDP-galactose in galactosyltransferase reaction, 2fold decrease in Km-value for lactose, 1.8fold increase in turnover number of hydrolase reaction, 2.3fold increase in Km-value for UDP-galactose in hydrolase reaction
W314Y
-
29fold decrease in turnover number of galactosyltransferase reaction, 1.67fold increase in Km-value for UDP-galactose in galactosyltransferase reaction, 2fold decrease in Km-value for lactose, 1.1fold increase in turnover number of hydrolase reaction, 1.4fold increase in Km-value for UDP-galactose in hydrolase reaction. Mutation does not affect the overall structure of the enzyme or its interactions with ligands
W356T
-
12.3fold decrease in turnover number of galactosyltransferase reaction, 2fold increase in Km-value for UDP-galactose in galactosyltransferase reaction, 6.3fold increase in Km-value for lactose, 14.5fold decrease in turnover number of hydrolase reaction, 2.5fold increase in Km-value for UDP-galactose in hydrolase reaction
N375V
-
exchange and truncation of terminal V376, 70% loss of activity
D102A
-
inactive
D104A
-
inactive
D116A
-
the mutant remains active
D118A
-
the mutant remains active
D28A
-
the mutant remains active
D30A
-
the mutant remains active
additional information