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Literature summary extracted from

  • Tsutsui, Y.; Ramakrishnan, B.; Qasba, P.K.
    Crystal structures of beta-1,4-galactosyltransferase 7 enzyme reveal conformational changes and substrate binding (2013), J. Biol. Chem., 288, 31963-31970.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.1.133 gene B4GALT7, recombinant expression of N-terminally His6-tagged enzyme Homo sapiens
2.4.1.133 gene B4GALT7, recombinant expression of N-terminally His6-tagged enzyme Drosophila melanogaster

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.4.1.133 hanging drop vapor diffusion method, using 100 mM imidazole (pH 6.5) and 750 mM sodium acetate or 100 mM HEPES buffer (pH 8.0), 100mM serinol, 1.0 M NaCl, 5% (w/v) PEG6000, and 10% 2,4-methanepentadiol Drosophila melanogaster
2.4.1.133 purified recombinant detagged enzyme in complex xylobiose, hanging drop vapor diffusion method, mixing of protein solution containing 10 mg/ml protein, 10 mM MnCl2, 10 mM UDP, and 10 mM xylobiose, with a reservoir solution containing 100mM HEPES, pH 8.0, 100mM serinol, 1.0 M NaCl, 5% PEG 6000, and 10% 2-methyl-2,4-pentanediol, crystals of enzyme mutants D318N and D211N protein in complex with UDP-Gal, by hanging drop vapor diffusion method, using a protein solution containing 10 mg/ml protein, 10 mM MnCl2, and 10 mM UDP-Gal, with the precipitating solution containing 100 mM HEPES, pH 8.0, 1.0 M NaCl, 5% PEG 6000, and 10% 2-methyl-2,4-pentanediol, X-ray diffraction structure determination and analysis Drosophila melanogaster
2.4.1.133 purified recombinant detagged wild-type and truncated enzymes, hanging drop vapor diffusion method, mixing of 15 mg/ml protein solution with reservoir solution containing 100 mM imidazole, pH 6.5, 5 mM MnCl2, and 750 mM sodium acetate, 18°C, X-ray diffraction structure determination and analysis, molecular replacement using the Drosophila melanogaster beta4GalT7 crystal structure, PDB ID 3LW6, as a search model. For crystal complexes, tetragonal crystals are soaked in the same pH 6.5 reservoir solution containing 15% 2-methyl-2,4-pentanediol at 18 °C for 20 h followed by a brief soak in 100 mM Tris, pH 8.0, 750 mM sodium acetate, 15% 2-methyl-2,4-pentanediol, 5 mM MnCl2, and 5 mM UDP-Gal at room temperature. The monoclinic crystals are obtained within 3 days using 100 mM Tris, pH 8.5, and 8% PEG 8000 as a reservoir solution by mixing it with 20 mg/ml beta4GalT7DELTA81 in 10 mM MES (pH 6.5), 150 mM NaCl, 5 mM MnCl2, and 5 mM UDP-galactose at 18 °C Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
2.4.1.133 D211N site-directed mutagenesis, inactive mutant Homo sapiens
2.4.1.133 D211N mutant with negligible catalytic activity Drosophila melanogaster
2.4.1.133 D211N site-directed mutagenesis, the catalytic base Asp211 is mutated to Asn residue, inactive mutant, substrate-bound crystal structure analysis Drosophila melanogaster
2.4.1.133 D318N mutant with negligible catalytic activity Drosophila melanogaster
2.4.1.133 additional information construction of truncated mutant beta4GalT7DELTA81 Homo sapiens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.4.1.38 Mn2+ required Homo sapiens
2.4.1.133 Mn2+ required Drosophila melanogaster
2.4.1.133 Mn2+ dependent on Drosophila melanogaster
2.4.1.133 Mn2+ dependent on, the manganese binding site is located at the N-terminal hinge region of the long loop, the conformational changes influence the manganese ion coordination, involving the Asp163-X-Asp165 and the His257-X-His259 motifs Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.1.38 UDP-alpha-D-galactose + N-acetyl-beta-D-glucosaminylglycopeptide Homo sapiens
-
 UDP + beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosaminylglycopeptide
-
 ?
2.4.1.133 UDP-alpha-D-galactose + O-beta-D-xylosyl-[protein] Homo sapiens
-
 UDP + 4-beta-D-galactosyl-O-beta-D-xylosyl-[protein]
-
 ?
2.4.1.133 UDP-alpha-D-galactose + O-beta-D-xylosyl-[protein] Drosophila melanogaster
-
 UDP + 4-beta-D-galactosyl-O-beta-D-xylosyl-[protein]
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.38 Homo sapiens Q9UBV7
-
-
2.4.1.133 Drosophila melanogaster Q9VBZ9
-
-
2.4.1.133 Drosophila melanogaster Q9VBZ9 gene B4GALT7
-
2.4.1.133 Homo sapiens Q9UBV7 gene B4GALT7
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.1.133 recombinant N-terminally His6-tagged enzyme by affinity chromatography, and cleavage of the tag by TEV protease Homo sapiens
2.4.1.133 recombinant N-terminally His6-tagged enzyme by affinity chromatography, and cleavage of the tag by TEV protease Drosophila melanogaster

Reaction

EC Number Reaction Comment Organism Reaction ID
2.4.1.133 UDP-alpha-D-galactose + [protein]-3-O-(beta-D-xylosyl)-L-serine = UDP + [protein]-3-O-(beta-D-galactosyl-(1->4)-beta-D-xylosyl)-L-serine catalytic mechanism, oveview Homo sapiens
a
2.4.1.133 UDP-alpha-D-galactose + [protein]-3-O-(beta-D-xylosyl)-L-serine = UDP + [protein]-3-O-(beta-D-galactosyl-(1->4)-beta-D-xylosyl)-L-serine catalytic mechanism, oveview Drosophila melanogaster
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.38 UDP-alpha-D-galactose + N-acetyl-beta-D-glucosaminylglycopeptide
-
 Homo sapiens UDP + beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosaminylglycopeptide
-
 ?
2.4.1.133 additional information donor and acceptor substrate binding, structure analysis, overview Drosophila melanogaster ?
-
 ?
2.4.1.133 additional information donor and acceptor substrate binding, strutcure analysis, overview Homo sapiens ?
-
 ?
2.4.1.133 UDP-alpha-D-galactose + O-beta-D-xylosyl-[protein]
-
 Homo sapiens UDP + 4-beta-D-galactosyl-O-beta-D-xylosyl-[protein]
-
 ?
2.4.1.133 UDP-alpha-D-galactose + O-beta-D-xylosyl-[protein]
-
 Drosophila melanogaster UDP + 4-beta-D-galactosyl-O-beta-D-xylosyl-[protein]
-
 ?
2.4.1.133 UDP-alpha-D-galactose + xylobiose
-
 Drosophila melanogaster UDP + ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.4.1.38 beta-1,4-GalT7
-
 Homo sapiens
2.4.1.38 beta1,4-galactosyltransferase 7
-
 Homo sapiens
2.4.1.133 beta-1,4-galactosyltransferase 7
-
 Homo sapiens
2.4.1.133 beta-1,4-galactosyltransferase 7
-
 Drosophila melanogaster
2.4.1.133 beta1,4-galactosyltransferase 7
-
 Drosophila melanogaster
2.4.1.133 beta4GalT7
-
 Homo sapiens
2.4.1.133 beta4GalT7
-
 Drosophila melanogaster

General Information

EC Number General Information Comment Organism
2.4.1.133 additional information the enzyme undergoes conformational changes upon substrate binding, human enzyme in its open and closed conformations, overview Homo sapiens
2.4.1.133 physiological function the enzyme is involved in proteoglycan synthesis Homo sapiens
2.4.1.133 physiological function the enzyme is involved in proteoglycan synthesis Drosophila melanogaster