Cloned (Comment) | Organism |
---|---|
gene M1 GAS SF370, DNA and amino acid sequence determination and analysis, cloning of the coding sequence for SpGH38 residues 1-901, overexpression as 3C protease cleavable N-terminal His6-tagged enzyme in Escherichia coli strain BL21(DE3) | Streptococcus pyogenes M1 GAS |
Crystallization (Comment) | Organism |
---|---|
purified recombinent enzyme free or in complex with the inhibitor swainsonine, sitting drop vapour diffusion method, mixing of 12 mg/ml protein in 100 mM Tris, pH 8.5, 1.5 M (NH4)2SO4 and 12% v/v glycerol, with reservoir solution, containing 3% v/v glycerol, 54% v/v Tacsimate, pH 7.0, and 2% v/v PEG 6000, also acting as the cryo-protectant, crystals of the swainsonine complex form are obtained by soaking SpGH38 crystals for ,16 h in mother liquor supplemented with 2 mM swainsonine, X-ray diffraction structure determination and analysis at 1.9 A and 2.6 A resolution, respectively | Streptococcus pyogenes M1 GAS |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
swainsonine | binding structure, overview | Streptococcus pyogenes M1 GAS |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3.6 | - |
4-methylumbelliferyl alpha-D-mannopyranoside | pH 6.8, 37°C, recombinant enzyme | Streptococcus pyogenes M1 GAS | |
27 | - |
3-O-alpha-D-mannopyranosyl-alpha-D-mannopyranose | pH 6.8, 37°C, recombinant enzyme | Streptococcus pyogenes M1 GAS |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
intracellular | SpGH38 may be intracellular, absence of any classical signal peptide | Streptococcus pyogenes M1 GAS | 5622 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | involved in catalysis | Streptococcus pyogenes M1 GAS |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
high mannose N-glycan + H2O | Streptococcus pyogenes M1 GAS | - |
? | - |
? | |
additional information | Streptococcus pyogenes M1 GAS | higher eukaryotic GH38 alpha-mannosides play a key role in the modification and diversification of hybrid N-glycans, processes with strong cellular links to cancer and autoimmune disease | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptococcus pyogenes M1 GAS | Q99YP5 | gene M1 GAS SF370 | - |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration | Streptococcus pyogenes M1 GAS |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
Man5GlcNAc3-[protein] + 2 H2O = Man3GlcNAc3-[protein] + 2 alpha-D-mannopyranose | reaction mechanism via glycosyl-enzyme intermediate, overview | Streptococcus pyogenes M1 GAS |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2,4-dinitrophenyl alpha-D-mannopyranoside + H2O | - |
Streptococcus pyogenes M1 GAS | 2,4-dinitrophenol + alpha-D-mannopyranose | - |
? | |
3-O-alpha-D-mannopyranosyl-alpha-D-mannopyranose + H2O | the enzyme is only active with the alpha-1,3-linked disaccharide | Streptococcus pyogenes M1 GAS | alpha-D-mannopyranose | - |
? | |
4-methylumbelliferyl alpha-D-mannopyranoside + H2O | - |
Streptococcus pyogenes M1 GAS | 4-methylumbelliferol + alpha-D-mannopyranose | - |
? | |
4-nitrophenyl alpha-D-mannopyranoside + H2O | - |
Streptococcus pyogenes M1 GAS | 4-nitrophenol + alpha-D-mannopyranose | - |
? | |
high mannose N-glycan + H2O | - |
Streptococcus pyogenes M1 GAS | ? | - |
? | |
additional information | higher eukaryotic GH38 alpha-mannosides play a key role in the modification and diversification of hybrid N-glycans, processes with strong cellular links to cancer and autoimmune disease | Streptococcus pyogenes M1 GAS | ? | - |
? | |
additional information | SpGH38 is an alpha-mannosidase with specificity for alpha-1,3 mannosidic linkages, active site structure, overview | Streptococcus pyogenes M1 GAS | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | structure comparisons, overview | Streptococcus pyogenes M1 GAS |
Synonyms | Comment | Organism |
---|---|---|
alpha-1,3 mannosidase | - |
Streptococcus pyogenes M1 GAS |
alpha-mannosidase | - |
Streptococcus pyogenes M1 GAS |
GH38 alpha-mannosidase II | - |
Streptococcus pyogenes M1 GAS |
GH38 enzyme | - |
Streptococcus pyogenes M1 GAS |
More | the enzyme belongs to the family GH38 of alpha-mannosidases | Streptococcus pyogenes M1 GAS |
SpGH38 | - |
Streptococcus pyogenes M1 GAS |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Streptococcus pyogenes M1 GAS |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.03 | - |
4-methylumbelliferyl alpha-D-mannopyranoside | pH 6.8, 37°C, recombinant enzyme | Streptococcus pyogenes M1 GAS | |
6.4 | - |
3-O-alpha-D-mannopyranosyl-alpha-D-mannopyranose | pH 6.8, 37°C, recombinant enzyme | Streptococcus pyogenes M1 GAS |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.8 | - |
assay at | Streptococcus pyogenes M1 GAS |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.018 | - |
swainsonine | - |
Streptococcus pyogenes M1 GAS |
General Information | Comment | Organism |
---|---|---|
physiological function | the enzyme is perhaps functioning in the subsequent degradation of extracellular host glycans following their initial digestion by secreted glycosidases | Streptococcus pyogenes |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.01 | - |
2,4-dinitrophenyl alpha-D-mannopyranoside | pH 6.8, 37°C, recombinant enzyme | Streptococcus pyogenes | |
0.24 | - |
3-O-alpha-D-mannopyranosyl-alpha-D-mannopyranose | pH 6.8, 37°C, recombinant enzyme | Streptococcus pyogenes |