Literature summary for 2.4.1.87 extracted from

Tumbale, P.; Jamaluddin, H.; Thiyagarajan, N.; Brew, K.; Acharya, K.R.
Structural basis of UDP-galactose binding by alpha-1,3-galactosyltransferase (alpha3GT): role of negative charge on aspartic acid 316 in structure and activity (2008), Biochemistry, 47, 8711-8718.

Crystallization (Commentary)

Crystallization (Comment)	Organism
crystal structure of the Glu317Gln mutant in complex with UDP-Gal is reported	Bos taurus
purified recombinant enzyme mutants with bound UDP-Gal and Mn2+, hanging drop vapour diffusion at 16°C, mixing of 0.001 ml protein solution containing 5 mg/ml mutant E317Q in 20 mM MES-NaOH buffer, pH 6.0, and 10% glycerol, containing 10 mM MnCl2 and 10 mM UDP-Gal with 0.001 ml of reservoir solution containing 10% PEG 6000, 0.1 M Tris-HCl, pH 8.0, and 8% MPD, for mutant D316N and D316E, 0.001 ml of 5 mg/ml protein in in 20 mM MES-NaOH buffer, pH 6.0, with 10% glycerol, 10 mM UDP-Gal, 10 mM MnCl2, and 10 mM N-acetyllactosamine, are mixed with 0.001 ml of a reservoir solution containing 5-10% PEG 6000, 0.1 M Tris-HCl, pH 8.0, and 5-14% MPD, X-ray diffraction structure determination and analysis at 1.77-2.2 A resolution	Bos taurus

Crystallization (Comment)

Organism

crystal structure of the Glu317Gln mutant in complex with UDP-Gal is reported

Bos taurus

purified recombinant enzyme mutants with bound UDP-Gal and Mn2+, hanging drop vapour diffusion at 16°C, mixing of 0.001 ml protein solution containing 5 mg/ml mutant E317Q in 20 mM MES-NaOH buffer, pH 6.0, and 10% glycerol, containing 10 mM MnCl2 and 10 mM UDP-Gal with 0.001 ml of reservoir solution containing 10% PEG 6000, 0.1 M Tris-HCl, pH 8.0, and 8% MPD, for mutant D316N and D316E, 0.001 ml of 5 mg/ml protein in in 20 mM MES-NaOH buffer, pH 6.0, with 10% glycerol, 10 mM UDP-Gal, 10 mM MnCl2, and 10 mM N-acetyllactosamine, are mixed with 0.001 ml of a reservoir solution containing 5-10% PEG 6000, 0.1 M Tris-HCl, pH 8.0, and 5-14% MPD, X-ray diffraction structure determination and analysis at 1.77-2.2 A resolution

Bos taurus

Protein Variants

Protein Variants	Comment	Organism
D316E	mutant show modest reduction in kcat and Km for lactose. Strucutural studies with mutant D316E show that the negative charge is crucial for catalytic activity and needed for its interaction with Arg202 for an active site structure that facilitates the binding of UDP-gal in a catalytically competent conformation	Bos taurus
D316E	site-directed mutagenesis, a catalytic domain mutant, crystal structure determination with bound UDP-Gal and Mn2+	Bos taurus
D316N	mutant is inactive. Strucutural studies with mutant D316N show that the negative charge is crucial for catalytic activity and needed for its interaction with Arg202 for an active site structure that facilitates the binding of UDP-gal in a catalytically competent conformation	Bos taurus
D316N	site-directed mutagenesis, a catalytic domain mutant, crystal structure determination with bound UDP-Gal and Mn2+	Bos taurus
E317Q	crystal structure of the complex of mutant E317Q with UDP-galactose exhibiting a bent configuration stabilized by interactions of the galactose with multiple residues in the enzyme including those in a highly conserved region (His315 to Ser318) is shown	Bos taurus
E317Q	site-directed mutagenesis, a catalytic domain mutant, crystal structure determination with bound UDP-Gal and Mn2+	Bos taurus
H315Q	mutant protein shows modest changes in kinetic parameters for lactose	Bos taurus
H315R	mutant shows a major losss in catalytic activity arising from a 500fold reduction in kcat	Bos taurus
H319A	mutant shows a 2fold reduction for kcat	Bos taurus
H319E	mutant is inactive	Bos taurus
H319Y	mutant is inactive	Bos taurus
S318A	mutant shows a 10fold reduction for kcat	Bos taurus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	steady-state kinetics	Bos taurus

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mn2+	binding structure at the catalytic domain, structure model, overview	Bos taurus

Organism

Organism	UniProt	Comment	Textmining
Bos taurus	-	-	-
Bos taurus	P14769	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	structural basis of UDP-galactose binding at the catalytic domain of alpha3GT, overview	Bos taurus	?	-	?

Synonyms

Synonyms	Comment	Organism
alpha-1,3-galactosyltransferase	-	Bos taurus
alpha3GT	-	Bos taurus