Cloned (Comment) | Organism |
---|---|
gene B4GALT7, recombinant expression of N-terminally His6-tagged enzyme | Homo sapiens |
gene B4GALT7, recombinant expression of N-terminally His6-tagged enzyme | Drosophila melanogaster |
Crystallization (Comment) | Organism |
---|---|
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 |
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 |
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 | Comment | Organism |
---|---|---|
D211N | site-directed mutagenesis, inactive mutant | Homo sapiens |
D211N | mutant with negligible catalytic activity | Drosophila melanogaster |
D211N | site-directed mutagenesis, the catalytic base Asp211 is mutated to Asn residue, inactive mutant, substrate-bound crystal structure analysis | Drosophila melanogaster |
D318N | mutant with negligible catalytic activity | Drosophila melanogaster |
additional information | construction of truncated mutant beta4GalT7DELTA81 | Homo sapiens |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | required | Drosophila melanogaster | |
Mn2+ | dependent on | Drosophila melanogaster | |
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 | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-alpha-D-galactose + O-beta-D-xylosyl-[protein] | Homo sapiens | - |
UDP + 4-beta-D-galactosyl-O-beta-D-xylosyl-[protein] | - |
? | |
UDP-alpha-D-galactose + O-beta-D-xylosyl-[protein] | Drosophila melanogaster | - |
UDP + 4-beta-D-galactosyl-O-beta-D-xylosyl-[protein] | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Drosophila melanogaster | Q9VBZ9 | - |
- |
Drosophila melanogaster | Q9VBZ9 | gene B4GALT7 | - |
Homo sapiens | Q9UBV7 | gene B4GALT7 | - |
Purification (Comment) | Organism |
---|---|
recombinant N-terminally His6-tagged enzyme by affinity chromatography, and cleavage of the tag by TEV protease | Homo sapiens |
recombinant N-terminally His6-tagged enzyme by affinity chromatography, and cleavage of the tag by TEV protease | Drosophila melanogaster |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
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 | |
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 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | donor and acceptor substrate binding, structure analysis, overview | Drosophila melanogaster | ? | - |
? | |
additional information | donor and acceptor substrate binding, strutcure analysis, overview | Homo sapiens | ? | - |
? | |
UDP-alpha-D-galactose + O-beta-D-xylosyl-[protein] | - |
Homo sapiens | UDP + 4-beta-D-galactosyl-O-beta-D-xylosyl-[protein] | - |
? | |
UDP-alpha-D-galactose + O-beta-D-xylosyl-[protein] | - |
Drosophila melanogaster | UDP + 4-beta-D-galactosyl-O-beta-D-xylosyl-[protein] | - |
? | |
UDP-alpha-D-galactose + xylobiose | - |
Drosophila melanogaster | UDP + ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
beta-1,4-galactosyltransferase 7 | - |
Homo sapiens |
beta-1,4-galactosyltransferase 7 | - |
Drosophila melanogaster |
beta1,4-galactosyltransferase 7 | - |
Drosophila melanogaster |
beta4GalT7 | - |
Homo sapiens |
beta4GalT7 | - |
Drosophila melanogaster |
General Information | Comment | Organism |
---|---|---|
additional information | the enzyme undergoes conformational changes upon substrate binding, human enzyme in its open and closed conformations, overview | Homo sapiens |
physiological function | the enzyme is involved in proteoglycan synthesis | Homo sapiens |
physiological function | the enzyme is involved in proteoglycan synthesis | Drosophila melanogaster |