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Literature summary extracted from

  • Suits, M.D.; Zhu, Y.; Taylor, E.J.; Walton, J.; Zechel, D.L.; Gilbert, H.J.; Davies, G.J.
    Structure and kinetic investigation of Streptococcus pyogenes family GH38 alpha-mannosidase (2010), PLoS ONE, 5, e9006.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.24 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
3.2.1.114 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 (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.24 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
3.2.1.114 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

EC Number Inhibitors Comment Organism Structure
3.2.1.24 swainsonine binding structure, overview Streptococcus pyogenes M1 GAS
3.2.1.114 swainsonine binding structure, overview Streptococcus pyogenes M1 GAS

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.24 3.6
-
 4-methylumbelliferyl alpha-D-mannopyranoside pH 6.8, 37°C, recombinant enzyme Streptococcus pyogenes M1 GAS
3.2.1.24 27
-
 3-O-alpha-D-mannopyranosyl-alpha-D-mannopyranose pH 6.8, 37°C, recombinant enzyme Streptococcus pyogenes M1 GAS
3.2.1.114 3.6
-
 4-methylumbelliferyl alpha-D-mannopyranoside pH 6.8, 37°C, recombinant enzyme Streptococcus pyogenes M1 GAS
3.2.1.114 27
-
 3-O-alpha-D-mannopyranosyl-alpha-D-mannopyranose pH 6.8, 37°C, recombinant enzyme Streptococcus pyogenes M1 GAS

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.2.1.24 intracellular SpGH38 may be intracellular, absence of any classical signal peptide Streptococcus pyogenes M1 GAS 5622
-
3.2.1.114 intracellular SpGH38 may be intracellular, absence of any classical signal peptide Streptococcus pyogenes M1 GAS 5622
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.24 Zn2+ involved in catalysis Streptococcus pyogenes M1 GAS
3.2.1.114 Zn2+ involved in catalysis Streptococcus pyogenes M1 GAS

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.24 high mannose N-glycan + H2O Streptococcus pyogenes M1 GAS
-
 ?
-
 ?
3.2.1.24 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 ?
-
 ?
3.2.1.114 high mannose N-glycan + H2O Streptococcus pyogenes M1 GAS
-
 ?
-
 ?
3.2.1.114 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

EC Number Organism UniProt Comment Textmining
3.2.1.24 Streptococcus pyogenes M1 GAS Q99YP5
-
-
3.2.1.114 Streptococcus pyogenes M1 GAS Q99YP5 gene M1 GAS SF370
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.24 recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration Streptococcus pyogenes M1 GAS
3.2.1.114 recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration Streptococcus pyogenes M1 GAS

Reaction

EC Number Reaction Comment Organism Reaction ID
3.2.1.114 Man5GlcNAc3-[protein] + 2 H2O = Man3GlcNAc3-[protein] + 2 alpha-D-mannopyranose reaction mechanism via glycosyl-enzyme intermediate, overview Streptococcus pyogenes M1 GAS
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.24 2,4-dinitrophenyl alpha-D-mannopyranoside + H2O
-
 Streptococcus pyogenes M1 GAS 2,4-dinitrophenol + alpha-D-mannopyranose
-
 ?
3.2.1.24 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
-
 ?
3.2.1.24 4-methylumbelliferyl alpha-D-mannopyranoside + H2O
-
 Streptococcus pyogenes M1 GAS 4-methylumbelliferol + alpha-D-mannopyranose
-
 ?
3.2.1.24 4-nitrophenyl alpha-D-mannopyranoside + H2O
-
 Streptococcus pyogenes M1 GAS 4-nitrophenol + alpha-D-mannopyranose
-
 ?
3.2.1.24 high mannose N-glycan + H2O
-
 Streptococcus pyogenes M1 GAS ?
-
 ?
3.2.1.24 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 ?
-
 ?
3.2.1.24 additional information SpGH38 is an alpha-mannosidase with specificity for alpha-1,3 mannosidic linkages, active site structure, overview Streptococcus pyogenes M1 GAS ?
-
 ?
3.2.1.114 2,4-dinitrophenyl alpha-D-mannopyranoside + H2O
-
 Streptococcus pyogenes M1 GAS 2,4-dinitrophenol + alpha-D-mannopyranose
-
 ?
3.2.1.114 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
-
 ?
3.2.1.114 4-methylumbelliferyl alpha-D-mannopyranoside + H2O
-
 Streptococcus pyogenes M1 GAS 4-methylumbelliferol + alpha-D-mannopyranose
-
 ?
3.2.1.114 4-nitrophenyl alpha-D-mannopyranoside + H2O
-
 Streptococcus pyogenes M1 GAS 4-nitrophenol + alpha-D-mannopyranose
-
 ?
3.2.1.114 high mannose N-glycan + H2O
-
 Streptococcus pyogenes M1 GAS ?
-
 ?
3.2.1.114 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 ?
-
 ?
3.2.1.114 additional information SpGH38 is an alpha-mannosidase with specificity for alpha-1,3 mannosidic linkages, active site structure, overview Streptococcus pyogenes M1 GAS ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.24 More structure comparisons, overview Streptococcus pyogenes M1 GAS
3.2.1.114 More structure comparisons, overview Streptococcus pyogenes M1 GAS

Synonyms

EC Number Synonyms Comment Organism
3.2.1.24 alpha-1,3 mannosidase
-
 Streptococcus pyogenes M1 GAS
3.2.1.24 alpha-mannosidase
-
 Streptococcus pyogenes M1 GAS
3.2.1.24 GH38 alpha-mannosidase II
-
 Streptococcus pyogenes M1 GAS
3.2.1.24 GH38 enzyme
-
 Streptococcus pyogenes M1 GAS
3.2.1.24 More the enzyme belongs to the family GH38 of alpha-mannosidases Streptococcus pyogenes M1 GAS
3.2.1.24 SpGH38
-
 Streptococcus pyogenes M1 GAS
3.2.1.114 alpha-1,3 mannosidase
-
 Streptococcus pyogenes M1 GAS
3.2.1.114 alpha-mannosidase
-
 Streptococcus pyogenes M1 GAS
3.2.1.114 GH38 alpha-mannosidase II
-
 Streptococcus pyogenes M1 GAS
3.2.1.114 GH38 enzyme
-
 Streptococcus pyogenes M1 GAS
3.2.1.114 More the enzyme belongs to the family GH38 of alpha-mannosidases Streptococcus pyogenes M1 GAS
3.2.1.114 SpGH38
-
 Streptococcus pyogenes M1 GAS

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.24 37
-
 assay at Streptococcus pyogenes M1 GAS
3.2.1.114 37
-
 assay at Streptococcus pyogenes M1 GAS

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.24 0.03
-
 4-methylumbelliferyl alpha-D-mannopyranoside pH 6.8, 37°C, recombinant enzyme Streptococcus pyogenes M1 GAS
3.2.1.24 6.4
-
 3-O-alpha-D-mannopyranosyl-alpha-D-mannopyranose pH 6.8, 37°C, recombinant enzyme Streptococcus pyogenes M1 GAS
3.2.1.114 0.03
-
 4-methylumbelliferyl alpha-D-mannopyranoside pH 6.8, 37°C, recombinant enzyme Streptococcus pyogenes M1 GAS
3.2.1.114 6.4
-
 3-O-alpha-D-mannopyranosyl-alpha-D-mannopyranose pH 6.8, 37°C, recombinant enzyme Streptococcus pyogenes M1 GAS

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.24 6.8
-
 assay at Streptococcus pyogenes M1 GAS
3.2.1.114 6.8
-
 assay at Streptococcus pyogenes M1 GAS

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.2.1.24 0.018
-
 swainsonine pH 6.8, 37°C Streptococcus pyogenes M1 GAS
3.2.1.114 0.018
-
 swainsonine
-
 Streptococcus pyogenes M1 GAS

General Information

EC Number General Information Comment Organism
3.2.1.24 physiological function the enzyme is perhaps functioning in the subsequent degradation of extracellular host glycans following their initial digestion by secreted glycosidases Streptococcus pyogenes M1 GAS
3.2.1.114 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 [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.24 0.01
-
 2,4-dinitrophenyl alpha-D-mannopyranoside pH 6.8, 37°C, recombinant enzyme Streptococcus pyogenes M1 GAS
3.2.1.24 0.24
-
 3-O-alpha-D-mannopyranosyl-alpha-D-mannopyranose pH 6.8, 37°C, recombinant enzyme Streptococcus pyogenes M1 GAS
3.2.1.114 0.01
-
 2,4-dinitrophenyl alpha-D-mannopyranoside pH 6.8, 37°C, recombinant enzyme Streptococcus pyogenes
3.2.1.114 0.24
-
 3-O-alpha-D-mannopyranosyl-alpha-D-mannopyranose pH 6.8, 37°C, recombinant enzyme Streptococcus pyogenes