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Literature summary for 2.4.1.90 extracted from

  • Ramasamy, V.; Ramakrishnan, B.; Boeggeman, E.; Qasba, P.K.
    The role of tryptophan 314 in the conformational changes of beta1,4-galactosyltransferase-I (2003), J. Mol. Biol., 331, 1065-1076.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
alpha-lactalbumin enzyme binds to the effector Mus musculus

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type enzyme and mutant W314A in Escherichia coli in inclusion bodies Mus musculus

Crystallization (Commentary)

Crystallization (Comment) Organism
catalytic enzyme domain without complexed substrate, conf-I' conformation, X-ray diffraction structure determination and analysis at 1.9 A resolution Bos taurus
W314A enzyme mutant complexed with alpha-lactalbumin in presence of N-acetylglucoamine, UDP, and Mn2+, conf-II conformation, X-ray diffraction structure determination and analysis at 2.3 A resolution Mus musculus

Protein Variants

Protein Variants Comment Organism
W314A site-directed mutagenesis, mutant shows highly reduced activity, i.e. 0.6% of wild-type galactosyltransferase activity, substrate binding, and reduced binding to the effector alpha-lactalbumin as well as reduced susceptibility to cleavage by proteases Glu-C and Lys-C compared to the wild-type enzyme, overview Mus musculus

Localization

Localization Comment Organism GeneOntology No. Textmining
Golgi apparatus
-
Mus musculus 5794
-
Golgi apparatus
-
Bos taurus 5794
-
membrane
-
Mus musculus 16020
-
membrane
-
Bos taurus 16020
-
additional information enzyme possesses a membrane-spanning domain, a cytoplasmic domain, a stem region, and the catalytic domain that faces the lumen Mus musculus
-
-
additional information enzyme possesses a membrane-spanning domain, a cytoplasmic domain, a stem region, and the catalytic domain that faces the lumen Bos taurus
-
-

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ required by full length wild-type enzyme Bos taurus
Mn2+ required by full length wild-type enzyme, coordination structure Mus musculus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
UDP-galactose + N-acetyl-D-glucosamine Mus musculus
-
UDP + N-acetyllactosamine
-
?
UDP-galactose + N-acetyl-D-glucosamine Bos taurus
-
UDP + N-acetyllactosamine
-
?

Organism

Organism UniProt Comment Textmining
Bos taurus
-
-
-
Mus musculus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type enzyme and mutant W314A from Escherichia coli after solubilization from inclusion bodies by alpha-lactalbumin affinity chromatography Mus musculus

Reaction

Reaction Comment Organism Reaction ID
UDP-alpha-D-galactose + N-acetyl-D-glucosamine = UDP + N-acetyllactosamine catalytic mechanism, enzyme undergoes critical conformational changes upon substrate binding from an open, conf-I, to a closed conformation, conf-II, involving 2 flexible loops, a small and a long one, with Trp314 of the small loop being critical for the change interacting with both substrates, catalytic site structure Bos taurus
UDP-alpha-D-galactose + N-acetyl-D-glucosamine = UDP + N-acetyllactosamine catalytic mechanism, enzyme undergoes critical conformational changes upon substrate binding from an open, conf-I, to a closed conformation, conf-II, involving 2 flexible loops, a small and a long one, with Trp314 of the small loop being critical for the change interacting with both substrates, catalytic site structure, substrate binding structure Mus musculus

Renatured (Commentary)

Renatured (Comment) Organism
refolding of the wild-type enzyme and mutant W314A from inclusion bodies after recombinant expression in Escherichia coli Mus musculus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
UDP-galactose + N-acetyl-D-glucosamine
-
Mus musculus UDP + N-acetyllactosamine
-
?
UDP-galactose + N-acetyl-D-glucosamine
-
Bos taurus UDP + N-acetyllactosamine
-
?
UDP-galactose + N-acetyl-D-glucosamine substrate binding structure Mus musculus UDP + N-acetyllactosamine
-
?

Subunits

Subunits Comment Organism
More enzyme possesses a membrane-spanning domain, a cytoplasmic domain, a stem region, and the catalytic domain that faces the lumen Mus musculus
More enzyme possesses a membrane-spanning domain, a cytoplasmic domain, a stem region, and the catalytic domain that faces the lumen Bos taurus

Synonyms

Synonyms Comment Organism
beta1,4-galactosyltransferase-I
-
Mus musculus
beta1,4-galactosyltransferase-I
-
Bos taurus
beta4Gal-T1
-
Mus musculus
beta4Gal-T1
-
Bos taurus